U.S. patent number 7,208,500 [Application Number 10/928,674] was granted by the patent office on 2007-04-24 for thienopyridine-phenylacetamides and their derivatives useful as new anti-angiogenic agents.
This patent grant is currently assigned to Agouron Pharmaceuticals, Inc.. Invention is credited to Mingying He, Robert Steven Kania, Jihong Lou.
United States Patent |
7,208,500 |
Lou , et al. |
April 24, 2007 |
Thienopyridine-phenylacetamides and their derivatives useful as new
anti-angiogenic agents
Abstract
The invention relates to compounds represented by Formula (I):
##STR00001## and to prodrugs thereof, pharmaceutically acceptable
salts or solvates of said compounds or said prodrugs, wherein each
of X.sup.1 X.sup.5 and R.sup.1 R.sup.5 are defined herein. The
invention also relates to pharmaceutical compositions containing
the compounds of Formula (I) and to methods of treating
hyperproliferative disorders in a mammal by administering compounds
of Formula (I).
Inventors: |
Lou; Jihong (San Diego, CA),
Kania; Robert Steven (San Diego, CA), He; Mingying (San
Diego, CA) |
Assignee: |
Agouron Pharmaceuticals, Inc.
(San Diego, CA)
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Family
ID: |
34272772 |
Appl.
No.: |
10/928,674 |
Filed: |
August 26, 2004 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20050090509 A1 |
Apr 28, 2005 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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60499077 |
Aug 29, 2003 |
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Current U.S.
Class: |
514/301;
546/301 |
Current CPC
Class: |
A61P
35/00 (20180101); A61P 43/00 (20180101); A61P
3/10 (20180101); C07D 495/04 (20130101); A61P
9/10 (20180101); A61P 27/02 (20180101); A61P
9/00 (20180101) |
Current International
Class: |
C07D
409/02 (20060101); A61K 31/381 (20060101) |
Field of
Search: |
;546/114 ;514/301 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
|
|
|
|
|
606046 |
|
Oct 1997 |
|
EP |
|
1086705 |
|
Mar 2001 |
|
EP |
|
931788 |
|
Nov 2002 |
|
EP |
|
WO90/05719 |
|
May 1990 |
|
WO |
|
WO95/19970 |
|
Jul 1995 |
|
WO |
|
WO95/21613 |
|
Aug 1995 |
|
WO |
|
WO95/23141 |
|
Aug 1995 |
|
WO |
|
WO96/14843 |
|
May 1996 |
|
WO |
|
WO96/27583 |
|
Sep 1996 |
|
WO |
|
WO96/30347 |
|
Oct 1996 |
|
WO |
|
WO96/33172 |
|
Oct 1996 |
|
WO |
|
WO96/40142 |
|
Dec 1996 |
|
WO |
|
WO97/13760 |
|
Apr 1997 |
|
WO |
|
WO97/13771 |
|
Apr 1997 |
|
WO |
|
WO97/22596 |
|
Jun 1997 |
|
WO |
|
WO97/32856 |
|
Sep 1997 |
|
WO |
|
WO97/34876 |
|
Sep 1997 |
|
WO |
|
WO97/49688 |
|
Dec 1997 |
|
WO |
|
WO98/02434 |
|
Jan 1998 |
|
WO |
|
WO98/02437 |
|
Jan 1998 |
|
WO |
|
WO98/02438 |
|
Jan 1998 |
|
WO |
|
WO98/03516 |
|
Jan 1998 |
|
WO |
|
WO98/07697 |
|
Feb 1998 |
|
WO |
|
WO98/14451 |
|
Apr 1998 |
|
WO |
|
WO98/23613 |
|
Jun 1998 |
|
WO |
|
WO98/30566 |
|
Jul 1998 |
|
WO |
|
WO98/33768 |
|
Aug 1998 |
|
WO |
|
WO98/34915 |
|
Aug 1998 |
|
WO |
|
WO98/34918 |
|
Aug 1998 |
|
WO |
|
WO98/51344 |
|
Nov 1998 |
|
WO |
|
WO98/54093 |
|
Dec 1998 |
|
WO |
|
WO99/10349 |
|
Mar 1999 |
|
WO |
|
WO99/16755 |
|
Apr 1999 |
|
WO |
|
WO99/24440 |
|
May 1999 |
|
WO |
|
WO99/35132 |
|
Jul 1999 |
|
WO |
|
WO99/35146 |
|
Jul 1999 |
|
WO |
|
WO 99/46268 |
|
Sep 1999 |
|
WO |
|
WO99/52889 |
|
Oct 1999 |
|
WO |
|
WO99/52910 |
|
Oct 1999 |
|
WO |
|
WO99/61422 |
|
Dec 1999 |
|
WO |
|
WO 00/00493 |
|
Jan 2000 |
|
WO |
|
WO00/16781 |
|
Mar 2000 |
|
WO |
|
WO00/37107 |
|
Jun 2000 |
|
WO |
|
WO00/38665 |
|
Jul 2000 |
|
WO |
|
WO00/38715 |
|
Jul 2000 |
|
WO |
|
WO00/38716 |
|
Jul 2000 |
|
WO |
|
WO00/38717 |
|
Jul 2000 |
|
WO |
|
WO00/38718 |
|
Jul 2000 |
|
WO |
|
WO00/38719 |
|
Jul 2000 |
|
WO |
|
WO00/38730 |
|
Jul 2000 |
|
WO |
|
WO00/38786 |
|
Jul 2000 |
|
WO |
|
WO01/16306 |
|
Mar 2001 |
|
WO |
|
WO01/21771 |
|
Mar 2001 |
|
WO |
|
WO01/70268 |
|
Sep 2001 |
|
WO |
|
WO01/74296 |
|
Oct 2001 |
|
WO |
|
WO01/74360 |
|
Oct 2001 |
|
WO |
|
WO01/85796 |
|
Nov 2001 |
|
WO |
|
WO02/30453 |
|
Apr 2002 |
|
WO |
|
WO02/41882 |
|
May 2002 |
|
WO |
|
WO02/064170 |
|
Aug 2002 |
|
WO |
|
WO02/070494 |
|
Sep 2002 |
|
WO |
|
WO03/006059 |
|
Jan 2003 |
|
WO |
|
WO 03/009852 |
|
Feb 2003 |
|
WO |
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WO03/035047 |
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May 2003 |
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WO |
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Other References
Parikh et al., Expression and Regulation of the Novel Vascular
Endothelial Growth Factor Receptor Neuropilin-1 by Epidermal Growth
Factor in Human Pancreatic Carcinoma, Cancer, 98(4):720-729 (2003).
cited by examiner .
Sumitomo et al., Immunohistochemical study of fibroblast Growth
Factor-2 (FGF-2) and Fibroblast Growth Factor Receptor (FGF-R) in
experimental squamous Cell Carcinoma of Rat Submandibular Gland,
Oral Oncology 35:98-104 (1999). cited by examiner .
Zhu & Witte, Inhibition of Tumor Growth and Metastasis by
Targeting Tumor-Associated Angiogenesis with Antagonists to the
Receptors of Vascular Endothelial Growth Factor, Investigational
New Drugs, 17:195-212 (1999). cited by examiner.
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Primary Examiner: Chang; Celia
Assistant Examiner: Balls; R. James
Attorney, Agent or Firm: Pugmire; Matthew J. Zielinski;
Bryan C.
Parent Case Text
This application claims the benefit of U.S. Provisional Application
No. 60/499,077, filed Aug. 29, 2003, the disclosure of which is
incorporated by reference herein in its entirety.
Claims
What is claimed is:
1. A compound represented by Formula (I): ##STR00163## wherein (a)
X.sup.1 is O or CR.sup.2aR.sup.2b; (b) X.sup.2 is CR.sup.1c; (c)
X.sup.3 is CR.sup.1d; (d) X.sup.4 is O; (e) X.sup.5 is CR.sup.4c;
each of R.sup.1a, R.sup.1b, R.sup.1c, and R.sup.1d is independently
selected from the group consisting of hydrogen, halogen, (C.sub.1
C.sub.6)alkoxy, (C.sub.1 C.sub.6)alkyl, (C.sub.1
C.sub.6)fluoroalkoxy, and (C.sub.1 C.sub.6)fluoroalkyl; (g) each of
R.sup.2a and R.sup.2b is independently selected from H, halogen, or
a moiety, (optionally substituted with 1 to 3 independently
selected Y.sup.1 groups) selected from the group consisting of
(C.sub.1 C.sub.6)alkoxy, (C.sub.1 C.sub.6)alkylamine and (C.sub.1
C.sub.6)alkyl, wherein any number of the hydrogen atoms on the
(C.sub.1 C.sub.6)alkoxy and (C.sub.1 C.sub.6)alkyl groups may be
optionally replaced with F; or R.sup.2a and R.sup.2b together can
be oxo or a moiety (optionally substituted with 1 to 3
independently selected Y.sup.1 groups) selected from the group
consisting of (C.sub.3 C.sub.6)cycloalkyl, 3 6 membered
heterocycloalkyl and .dbd.CH--(C.sub.1 C.sub.5)alkyl; (h) R.sup.3
is H or a moiety (optionally substituted with 1 3 independently
selected Y.sup.2 groups) selected from the group consisting of
--(CZ.sup.1Z.sup.2).sub.sCN, --(CZ.sup.1Z.sup.2).sub.s--(C.sub.3
C.sub.8)cycloalkyl, --(CZ.sup.1Z.sup.2).sub.s--(C.sub.5
C.sub.8)cycloalkenyl, (C.sub.2 C.sub.6)alkenyl, (C.sub.2
C.sub.6)alkynyl, --(CZ.sup.1Z.sup.2).sub.s-aryl,
--(CZ.sup.1Z.sup.2).sub.s-heterocycle, and (C.sub.1 C.sub.8)alkyl,
where s is 0, 1, 2, or 3, and wherein when s is 2 or 3, the
--CZ.sup.1Z.sup.2 units may be the same or different; (i) each of
R.sup.4a, R.sup.4b, and R.sup.4c is independently selected from the
group consisting of H, F, Cl, CF.sub.3, CH.sub.3, OCH.sub.3, and
OCF.sub.3; (j) R.sup.5 is selected from the group consisting of
hydrogen, nitro, halogen, azido, --NR.sup.6aR.sup.6b,
--NR.sup.6aSO.sub.2R.sup.6b, --NR.sup.6aC(O)R.sup.6b,
--OC(O)R.sup.6b, --NR.sup.6aC(O)OR.sup.6b,
--OC(O)NR.sup.6aR.sup.6b, --OR.sup.6a, --SR.sup.6a, --S(O)R.sup.6a,
--SO.sub.2R.sup.6a, --SO.sub.3R.sup.6a,
--SO.sub.2NR.sup.6aR.sup.6b, --COR.sup.6a, --CO.sub.2R.sup.6b,
--CONR.sup.6aR.sup.6b, --(C.sub.1 C.sub.4)fluoroalkyl, --(C.sub.1
C.sub.4)fluoroalkoxy, --(CZ.sup.3Z.sup.4).sub.tCN, and a moiety
selected from the group consisting of
--(CZ.sup.3Z.sup.4).sub.t-aryl,
--(CZ.sup.3Z.sup.4).sub.t-heterocycle, (C.sub.2 C.sub.6)alkynyl,
--(CZ.sup.3Z.sup.4).sub.t--(C.sub.3 C.sub.6)cycloalkyl,
--(CZ.sup.3Z.sup.4).sub.t--(C.sub.5 C.sub.6)cycloalkenyl, (C.sub.2
C.sub.6)alkenyl, and (C.sub.1 C.sub.6)alkyl, which is optionally
substituted with 1 to 3 independently selected Y.sup.2 groups,
where t is 0, 1, 2, or 3, and wherein when t is 2 or 3, the
CZ.sup.3Z.sup.4 units may be the same or different; (k) each
R.sup.6a and R.sup.6b is independently selected from the group
consisting of hydrogen and a moiety selected from the group
consisting of --(CZ.sup.5Z.sup.6).sub.u--(C.sub.3
C.sub.6)cycloalkyl, --(CZ.sup.5Z.sup.6).sub.u--(C.sub.5
C.sub.6)cycloalkenyl, --(CZ.sup.5Z.sup.6).sub.u-aryl,
--(CZ.sup.5Z.sup.6).sub.u-heterocycle, (C.sub.2 C.sub.6)alkenyl,
and (C.sub.1 C.sub.6)alkyl, which is optionally substituted with 1
to 3 independently selected Y.sup.3 groups, where u is 0, 1, 2, or
3, and wherein when u is 2 or 3, the CZ.sup.5Z.sup.6 units may be
the same or different, or R.sup.6a and R.sup.6b taken together can
with adjacent atoms form a heterocycle; (l) each Z.sup.1, Z.sup.2,
Z.sup.3, Z.sup.4, Z.sup.5, and Z.sup.6 is independently selected
from the group consisting of H, F, and (C.sub.1 C.sub.6)alkyl, or
each Z.sup.1 and Z.sup.2, Z.sup.3 and Z.sup.4, or Z.sup.5 and
Z.sup.6 are selected together to form a carbocycle, or two Z.sup.1,
Z.sup.3 or Z.sup.3 groups on adjacent carbon atoms are selected
together to optionally form a carbocycle; and (m) each Y.sup.1 is
independently selected from the group consisting of halogen, cyano,
nitro, azido, --OH, --NH.sub.2, (C.sub.1 C.sub.6)alkoxy, (C.sub.1
C.sub.6)alkylamino, (C.sub.1 C.sub.6)dialkylamino, (C.sub.1
C.sub.6)alkyl, (C.sub.2 C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl,
(C.sub.1 C.sub.6)haloalkyl, (C.sub.1 C.sub.6)haloalkoxy, --(C.sub.3
C.sub.6)cycloalkyl; (n) each Y.sup.2 and Y.sup.3 is independently
selected and (i) is selected from the group consisting of halogen,
cyano, nitro, tetrazolyl, guanidino, amidino, methylguanidino,
azido, --C(O)Z.sup.7, --OC(O)NH.sub.2, --OC(O)NHZ.sup.7,
--OC(O)NZ.sup.7Z.sup.8, --NHC(O)Z.sup.7, --NHC(O)NH.sub.2,
--NHC(O)NHZ.sup.7, --NHC(O)NZ.sup.7Z.sup.8, --C(O)OH,
--C(O)OZ.sup.7, --C(O)NH.sub.2, --C(O)NHZ.sup.7,
--C(O)NZ.sup.7Z.sup.8, --P(O).sub.3H.sub.2,
--P(O).sub.3(Z.sup.7).sub.2, --S(O).sub.3H, --S(O)Z.sup.7,
--S(O).sub.2Z.sup.7, --S(O).sub.3Z.sup.7, -Z.sup.7, --OZ.sup.7,
--OH, --NH.sub.2, --NHZ.sup.7, --NZ.sup.7Z.sup.8,
--C(.dbd.NH)NH.sub.2, --C(.dbd.NOH)NH.sub.2, --N-morpholino,
(C.sub.2 C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl, (C.sub.1
C.sub.6)haloalkyl, (C.sub.2 C.sub.6)haloalkenyl, (C.sub.2
C.sub.6)haloalkynyl, (C.sub.1 C.sub.6)haloalkoxy,
--(CZ.sup.9Z.sup.10).sub.rNH.sub.2,
--(CZ.sup.9Z.sup.10).sub.rNHZ.sup.3,
--(CZ.sup.9Z.sup.10).sub.rNZ.sup.7Z.sup.8,
--X.sup.6(CZ.sup.9Z.sup.10).sub.r--(C.sub.3 C.sub.8) cycloalkyl,
--X.sup.6(CZ.sup.9Z.sup.10).sub.r--(C.sub.5 C.sub.8)cycloalkenyl,
--X.sup.6(CZ.sup.9Z.sup.10).sub.r-aryl, and
--X.sup.6(CZ.sup.9Z.sup.10).sub.r-heterocycle; r is 1, 2, 3, or 4;
X.sup.6 is O, S, NH, --C(O)--, --C(O)NH--, --C(O)O--, --S(O)--,
--S(O).sub.2--, or --S(O).sub.3--; Z.sup.7 and Z.sup.8 are
independently selected from the group consisting of alkyl of 1 to
12 carbon atoms, alkenyl of 2 to 12 carbon atoms, alkynyl of 2 to
12 carbon atoms, cycloalkyl of 3 to 8 carbon atoms, cycloalkenyl of
5 to 8 carbon atoms, aryl of 6 to 14 carbon atoms, heterocycle of 5
to 14 ring atoms, aralkyl of 7 to 15 carbon atoms, and
heteroaralkyl of 5 to 14 ring atoms, or Z.sup.7 and Z.sup.8
together may optionally form a heterocycle; and Z.sup.9 and
Z.sup.10 are independently selected from the group consisting of
hydrogen, fluorine, alkyl of 1 to 12 carbon atoms, aryl of 6 to 14
carbon atoms, heteroaryl of about 5 to 14 ring atoms, aralkyl of 7
to 15 carbon atoms, and heteroaralkyl of 5 to 14 ring atoms, or
Z.sup.9 and Z.sup.10 are selected together to form a carbocycle, or
two Z.sup.9 groups on adjacent carbon atoms are selected together
to form a carbocycle; or (ii) any two Y.sup.2 or Y.sup.3 groups
attached to adjacent carbon atoms may be selected together to be
--O[C(Z.sup.9)(Z.sup.10)].sub.rO-- or
--O[C(Z.sup.9)(Z.sup.10)].sub.r+1--; or (iii) any two Y.sup.2 or
Y.sup.3 groups attached to the same or adjacent carbon atoms may be
selected together to form a carbocycle or heterocycle; and wherein
any of the above-mentioned substituents comprising a CH.sub.3
(methyl), CH.sub.2 (methylene), or CH (methine) group which is not
attached to a halogen, SO or SO.sub.2 group or to a N, O or S atom
optionally bears on said group a substituent selected from hydroxy,
halogen, (C.sub.1 C.sub.4)alkyl, (C.sub.1 C.sub.4)alkoxy and
--N[(C.sub.1 C.sub.4)alkyl][(C.sub.1 C.sub.4)alkyl]; or a
pharmaceutically acceptable salt or solvate thereof.
2. The compound according to claim 1, wherein X.sup.1 is O.
3. The compound according to claim 2, wherein X.sup.5 is CH.
4. The compound according to claim 1, wherein X.sup.1 is
CH.sub.2.
5. The compound according to claim 1, wherein X.sup.5 is CH.
6. The compound according to claim 5, wherein X.sup.1 is
CH.sub.2.
7. The compound according to claim 6, wherein each of R.sup.1a,
R.sup.1b, R.sup.1c, and R.sup.1d is independently H, F, or Cl.
8. The compound according to claim 7, wherein each of R.sup.4a,
R.sup.4b, and R.sup.4c is independently selected from the group
consisting of H and F.
9. The compound according to claim 1, wherein R.sup.3 is a (C.sub.1
C.sub.8)alkyl, optionally substituted with 1 3 independently
selected Y.sup.2 groups.
10. The compound according to claim 8, wherein R.sup.3 is a
(C.sub.1 C.sub.8)alkyl, optionally substituted with 1 3
independently selected Y.sup.2 groups.
11. The compound according to claim 1, wherein R.sup.3 is a
heterocycle, optionally substituted with 1 3 independently selected
Y.sup.2 groups.
12. The compound according to claim 8, wherein R.sup.3 is a
heterocycle, optionally substituted with 1 3 independently selected
Y.sup.2 groups.
13. The compound according to claim 1, wherein R.sup.5 is
--CONR.sup.6aR.sup.6b.
14. The compound according to claim 1, wherein R.sup.5 is a
--(CZ.sup.3Z.sup.4).sub.t-heterocycle.
15. The compound according to claim 8, wherein R.sup.5 is
--CONR.sup.6aR.sup.6b.
16. The compound according to claim 8, wherein R.sup.5 is
--(CZ.sup.3Z.sup.4).sub.t-heterocycle.
17. The compound according to claim 1, wherein X.sup.1 is
CR.sup.2aR.sup.2b.
18. The compound according to claim 17, wherein R.sup.5 is
--C(O)NR.sup.6aR.sup.6b or
--(CZ.sup.3Z.sup.4).sub.t-heterocycle.
19. The compound according to claim 18, wherein R.sup.3 is (C.sub.1
C.sub.6)alkyl or --(CZ.sup.1Z.sup.2).sub.s heterocycle.
20. The compound according to claim 19, wherein R.sup.3 is
heteroaryl.
21. The compound according to claim 20, wherein R.sup.5 is
--C(O)NR.sup.6aR.sup.6b or heteroaryl.
22. The compound according to claim 21, wherein R is
--C(O)NR.sup.6aR.sup.6b wherein R.sup.6a and R.sup.6b taken with
the nitrogen atom form a heterocycle or R.sup.5 is imidazolye.
23. The compound according to claim 22, wherein R.sup.1a, R.sup.1b,
R.sup.1c, R.sup.1d, R.sup.2a, R.sup.2b, R.sup.4c, R.sup.4b and
R.sup.4c are hydrogen, F or Cl.
24. The compound according to claim 23, wherein R.sup.1b, R.sup.1c,
R.sup.1d, R.sup.2a, R.sup.2b, R.sup.4a, R.sup.4b and R.sup.4c are
hydrogen.
25. A compound selected from the group consisting of:
N-(4,6-Dimethyl-pyridin-2-yl)-2-{3-fluoro-4-[2-(1-methyl-
1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide;
2-{4-[2-((3R, 4R)-3,4-Dihydroxy-pyrrolidine-1-carbonyl)-thieno
3,2-b]pyridin-7-yloxy]-phenyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide;
2-{4-[2-((R)-3-Dimethylamino-pyrrolidine-1-carbonyl)-thieno
[3,2-b]pyridin-7-yloxy]-phenyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide;
7-{4-[(4,6-Dimethyl-pyridin-2-ylcarbamoyl)-methyl]-phenoxy}-thieno[3,2-b]-
pyridine-2-carboxylic acid dimethylamide;
N-(4,6-Dimethyl-pyridin-2-yl)-2-{4-[2-(1-methyl-
1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide;
N-(5-Chloro-pyridin-2-yl)-2-{4-[2-(1-methyl-1H-imidazol-
2-yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide;
N-(4,6-Dimethyl-pyridin-2-yl)-2-{4-[2-((R)-3-hydroxy-pyrrolidine-
1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide;
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-
7-yloxy]-phenyl}-N-isoquinolin-3-yl-acetamide;
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-
7-yloxy]-phenyl}-N-phenyl-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(4-
,6-dimethyl-pyridin-2-yl)-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(6-
-methyl-pyridin-2-yl)-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5-
-trifluoromethyl-pyridin-2-yl)-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5-
-chloro-pyridin-2-yl)-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5-
-bromo-pyridin-2-yl)-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-is-
oquinolin-3-yl-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phe-
nyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide;
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phe-
nyl}-N-(5-methyl-1H-pyrazol-3-yl)-acetamide; and Butyl-carbamic
acid
4-[2-(azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl
ester; or a pharmaceutically acceptable salt or solvate
thereof.
26. A compound selected from the group consisting of: ##STR00164##
##STR00165## ##STR00166## ##STR00167## or a pharmaceutically
acceptable salt or solvate thereof.
27. A pharmaceutical composition comprising a therapeutically
effective amount of the compound of claim 1 and a pharmaceutically
acceptable carrier.
28. A method of producing the compound of claim 1, wherein X.sup.1
is CR.sup.2aR.sup.2b, comprising: (a) reacting a carboxylic acid
having the structure ##STR00168## with a chlorinating agent; and
(b) reacting the resulting product with H.sub.2N--R.sup.3.
29. The method of claim 28, wherein the chlorinating agent is
selected from the group consisting of thionyl chloride, oxalyl
chloride, and chlorine.
30. The method of claim 28, wherein the carboxylic acid is produced
by a method comprising: (a) reacting a compound having the formula
##STR00169## with a compound having the formula ##STR00170## in the
presence of a base.
31. A method of producing the compound of claim 1, wherein X.sup.1
is O, comprising: (a) reacting a compound of formula ##STR00171##
with a carbonyl electrophile; and (b) reacting the resulting
product with H.sub.2N--R.sup.3.
32. The method of claim 31, wherein the carbonyl electrophile is
phosgene.
Description
FIELD OF THE INVENTION
This invention relates to novel thienopyridine-phenylacetamides and
derivatives thereof, including pharmaceutically acceptable
derivatives, such as salts, prodrugs, solvates, and metabolites.
The compounds of the present invention inhibit the activity of
receptor kinases such as VEGFR and PDGRF that are required for cell
growth and differentiation and angiogenesis. Particularly, the
compounds in this invention inhibit VEGFR/KDR and therefore are
useful for treatment of diseases and conditions that are associated
with VEGFR/KDR activity, e.g., cancer and ophthalmic diseases such
as age-related macular degeneration and diabetic retinopathy. This
invention also relates to a method of using such compounds in the
treatment of hyperproliferative diseases in mammals, especially
humans, and to pharmaceutical compositions containing such
compounds.
BACKGROUND
A cell may become cancerous by virtue of the transformation of a
portion of its DNA into an oncogene (i.e., a gene that upon
activation leads to the formation of malignant tumor cells). Many
oncogenes encode proteins that are aberrant tyrosine kinases
capable of causing cell transformation. Alternatively, the
overexpression of a normal proto-oncogenic tyrosine kinase may also
result in proliferative disorders, sometimes resulting in a
malignant phenotype.
Receptor tyrosine kinases are large enzymes that span the cell
membrane and possess an extracellular binding domain for growth
factors, a transmembrane domain, and an intracellular portion that
functions as a kinase to phosphorylate a specific tyrosine residue
in proteins and hence to influence cell proliferation. Tyrosine
kinases may be classified as growth factor receptor (e.g. EGFR,
PDGFR, FGFR and erbB2) or non-receptor (e.g. c-src and bcr-abl)
kinases. Such kinases may be aberrantly expressed in common human
cancers such as breast cancer, gastrointestinal cancers such as
colon, rectal or stomach cancer, leukemia, and ovarian, bronchial
or pancreatic cancer. Aberrant erbB2 activity has been implicated
in breast, ovarian, non-small cell lung, pancreatic, gastric and
colon cancers. Studies indicate that epidermal growth factor
receptor (EGFR) is mutated or overexpressed in many human cancers
such as brain, lung, squamous cell, bladder, gastric, breast, head
and neck, oesophageal, gynecological and thyroid cancers. Thus,
inhibitors of receptor tyrosine kinases may be useful as selective
inhibitors of the growth of mammalian cancer cells.
EGFR inhibitors may be useful in the treatment of pancreatitis and
kidney disease (such as proliferative glomerulonephritis and
diabetes-induced renal disease), and may reduce successful
blastocyte implantation and therefore may be useful as a
contraceptive. See PCT international application publication number
WO 95/19970 (published Jul. 27, 1995), hereby incorporated by
reference in its entirety.
Polypeptide growth factors, such as vascular endothelial growth
factor (VEGF) having a high affinity to the human kinase
insert-domain-containing receptor (KDR) or the murine fetal liver
kinase 1 (FLK-1) receptor have been associated with the
proliferation of endothelial cells and more particularly
vasculogenesis and angiogenesis. See PCT international application
publication number WO 95/21613 (published Aug. 17, 1995), hereby
incorporated by reference in its entirety. Agents that are capable
of binding to or modulating the KDR/FLK-1 receptor may be used to
treat disorders related to vasculogenesis or angiogenesis, such as
diabetes, diabetic retinopathy, age related macular degeneration,
hemangioma, glioma, melanoma, Kaposi's sarcoma and ovarian, breast,
lung, pancreatic, prostate, colon and epidermoid cancer.
Examples of compounds and methods that reportedly can be used to
treat hyperproliferative diseases are disclosed in the following
patents and applications: U.S. Pat. Nos. 6,534,524, 6,531,491 and
6,071,935; PCT international patent application publication nos. WO
00/38665 (published Jul. 6, 2001), WO 97/49688 (published Dec. 31,
1997), WO 98/23613 (published Jun. 4, 1998), WO 96/30347 (published
Oct. 3, 1996), WO 96/40142 (published Dec. 19, 1996), WO 97/13771
(published Apr. 17, 1997), WO 95/23141 (published Aug. 31, 1995),
WO 03/006059 (published Jan. 23, 2003), WO 03/035047 (published May
1, 2003), WO 02/064170 (published Aug. 22, 2002), WO 02/41882
(published May 30, 2002), WO 02/30453 (published Apr. 18, 2002), WO
01/85796 (published Nov. 15, 2001), WO 01/74360 (published Oct. 11,
2001), WO 01/74296 (published Oct. 11, 2001), WO 01/70268
(published Sep. 27, 2001) and WO 98/51344 (published Nov. 19,
1998); and European patent publication number EP 1086705 (published
Mar. 28, 2001). The foregoing patent and applications are each
incorporated herein by reference in their entirety.
SUMMARY
Described herein are compounds capable of modulating the activity
of receptor kinases such as VEGFR and PDGRF and methods for
utilizing such modulation in the treatment of cancer and other
proliferative disorders. Also described are compounds that mediate
and/or inhibit the activity of protein kinases, and pharmaceutical
compositions containing such compounds. Also described are
therapeutic or prophylactic use of such compounds and compositions,
and methods of treating cancer as well as other diseases associated
with unwanted angiogenesis and/or cellular proliferation, by
administering effective amounts of such compounds.
In one aspect are novel thienopyridine-phenylacetamide compounds.
In another aspect of the present invention are compounds that
modulate the activity of receptor kinases such as KDR/VEGFR2 kinase
in vitro and/or in vivo. According to a further aspect of the
present invention are compounds that can selectively modulate the
activity of receptor kinases such as KDR/VEGFR2 kinase. In yet
another aspect of the present invention, provided are
pharmaceutical compositions of such VEGFR2-modulating compounds,
including pharmaceutically acceptable prodrugs, pharmaceutically
acceptable solvates, pharmaceutically active metabolites, or
pharmaceutically acceptable salts thereof. According to yet another
aspect of the present invention, provided are syntheses schemes for
the preparation of such VEGFR2-modulating compounds, and
pharmaceutically acceptable prodrugs, pharmaceutically acceptable
solvates, pharmaceutically active metabolites, or pharmaceutically
acceptable salts thereof. In yet another aspect of the present
invention, methods are provided for modulating KDR/VEGFR2 kinase
which comprise contacting the VEGFR2-modulating compounds, or
pharmaceutically acceptable prodrugs, pharmaceutically acceptable
solvates, pharmaceutically active metabolites, or pharmaceutically
acceptable salts thereof, described herein, with KDR/VEGFR2 kinase.
In yet another aspect of the present invention, provided are
methods for treating patients comprising administering a
therapeutically effective amount of a VEGFR2-modulating compound,
or a pharmaceutically acceptable prodrug, pharmaceutically
acceptable solvate, pharmaceutically active metabolite, or
pharmaceutically acceptable salt thereof. In yet another aspect of
the present invention, are combination therapies involving
administration of an anti-neoplastic agent and an effective amount
of a VEGFR2-modulating compound, or a pharmaceutically acceptable
prodrug, pharmaceutically acceptable solvate, pharmaceutically
active metabolite, or pharmaceutically acceptable salt thereof.
In one aspect are compounds of Formula (I):
##STR00002## wherein (a) X.sup.1 is O or CR.sup.2aR.sup.2b. (b)
X.sup.2 is N or CR.sup.1c; (c) X.sup.3 is N or CR.sup.1d; (d)
X.sup.4 is O or S; (e) X.sup.5 is N or CR.sup.4c; (f) each of
R.sup.1a, R.sup.1b, R.sup.1c, and R.sup.1d is independently
selected from the group consisting of hydrogen, halogen, (C.sub.1
C.sub.6)alkoxy, (C.sub.1 C.sub.6)alkyl, (C.sub.1
C.sub.6)fluoroalkoxy, and (C.sub.1 C.sub.6)fluoroalkyl; (g) each of
R.sup.2a and R.sup.2b is independently selected from H, halogen, or
a moiety, optionally substituted with 1 to 3 independently selected
Y.sup.1 groups, selected from the group consisting of (C.sub.1
C.sub.6)alkoxy, (C.sub.1 C.sub.6)alkylamine and (C.sub.1
C.sub.6)alkyl, wherein any number of the hydrogen atoms on the
(C.sub.1 C.sub.6)alkoxy and (C.sub.1 C.sub.6)alkyl groups may be
optionally replaced with F; or R.sup.2a and R.sup.2b together can
be oxo or a moiety, optionally substituted with 1 to 3
independently selected Y.sup.1 groups, selected from the group
consisting of (C.sub.3 C.sub.6)cycloalkyl, 3 6 membered
heterocycloalkyl and .dbd.CH--(C.sub.1 to C.sub.5)alkyl; (h)
R.sup.3 is H or a moiety, optionally substituted with 1 3
independently selected Y.sup.2 groups, selected from the group
consisting of --(CZ.sup.1Z.sup.2).sub.sCN,
--(CZ.sup.1Z.sup.2).sub.s--(C.sub.3-C.sub.8)cycloalkyl,
--(CZ.sup.1Z.sup.2).sub.s--(C.sub.5-C.sub.8)cycloalkenyl, (C.sub.2
C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl,
--(CZ.sup.1Z.sup.2).sub.s-aryl,
--(CZ.sup.1Z.sup.2).sub.s-heterocycle, and (C.sub.1 C.sub.8)alkyl,
where s is 0, 1, 2, or 3, and wherein when s is 2 or 3, the
CZ.sup.1Z.sup.2 units may be the same or different; (i) each of
R.sup.4a, R.sup.4b, and R.sup.4c is independently selected from the
group consisting of H, F, Cl, CF.sub.3, CH.sub.3, OCH.sub.3, and
OCF.sub.3; (j) R.sup.5 is selected from the group consisting of
hydrogen, nitro, halogen, azido, --NR.sup.6aR.sup.6b,
--NR.sup.6aSO.sub.2R.sup.6b, --NR.sup.6aC(O)R.sup.6b,
--OC(O)R.sup.6b, --NR.sup.6aC(O)OR.sup.6b,
--OC(O)NR.sup.6aR.sup.6b, --OR.sup.6a, --SR.sup.a, --S(O)R.sup.6a,
--SO.sub.2R.sup.6a, --SO.sub.3R.sup.6a,
--SO.sub.2NR.sup.6aR.sub.6b, --COR.sup.6a, --CO.sub.2R.sup.6a,
--CONR.sup.6aR.sup.6b, --(C.sub.1 C.sub.4)fluoroalkyl, --(C.sub.1
C.sub.4)fluoroalkoxy, --(CZ.sup.3Z.sup.4).sub.tCN, and a moiety
selected from the group consisting of
--(CZ.sup.3Z.sup.4).sub.t-aryl,
--(CZ.sup.3Z.sup.4).sub.t-heterocycle, (C.sub.2 C.sub.6)alkynyl,
--(CZ.sup.3Z.sup.4).sub.t--(C.sub.3 C.sub.6)cycloalkyl,
--(CZ.sup.3Z.sup.4).sub.t--(C.sub.5 C.sub.6)cycloalkenyl, (C.sub.2
C.sub.6)alkenyl, and (C.sub.1 C.sub.6)alkyl, which is optionally
substituted with 1 to 3 independently selected Y.sup.2 groups,
where t is 0, 1, 2, or 3, and wherein when t is 2 or 3, the
CZ.sup.3Z.sup.4 units may be the same or different; (k) each
R.sup.6a and R.sup.6b is independently selected from the group
consisting of hydrogen and a moiety selected from the group
consisting of --(CZ.sup.5Z.sup.6).sub.u--(C.sub.3
C.sub.6)cycloalkyl, --(CZ.sup.5Z.sup.6).sub.u--(C.sub.5
C.sub.6)cycloalkenyl, --(CZ.sup.5Z.sup.6).sub.u-aryl,
--(CZ.sup.5Z.sup.6).sub.u-heterocycle, (C.sub.2 C.sub.6)alkenyl,
and (C.sub.1 C.sub.6)alkyl, which is optionally substituted with 1
to 3 independently selected Y.sup.3 groups, where u is 0, 1, 2, or
3, and wherein when u is 2 or 3, the CZ.sup.5Z.sup.6 units may be
the same or different, or R.sup.6a and R.sup.6b taken together can
with adjacent atoms form a heterocycle; (l) each Z.sup.1, Z.sup.2,
Z.sup.3, Z.sup.4, Z.sup.5, and Z.sup.6 is independently selected
from the group consisting of H, F, and (C.sub.1 C.sub.6)alkyl, or
each Z.sup.1 and Z.sup.2, Z.sup.3 and Z.sup.4, or Z.sup.5 and
Z.sup.6 are selected together to form a carbocycle, or two Z.sup.1,
Z.sup.3 or Z.sup.3 groups on adjacent carbon atoms are selected
together to optionally form a carbocycle; and (m) each Y.sup.1 is
independently selected from the group consisting of halogen, cyano,
nitro, azido, --OH, --NH.sub.2, (C.sub.1 C.sub.6)alkoxy, (C.sub.1
C.sub.6)alkylamino, (C.sub.1 C.sub.6)dialkylamino, (C.sub.1
C.sub.6)alkyl, (C.sub.2 C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl,
(C.sub.1 C.sub.6)haloalkyl, (C.sub.1 C.sub.6)haloalkoxy, --(C.sub.3
C.sub.6)cycloalkyl; (n) each Y.sup.2 and Y.sup.3 is independently
selected and (i) is selected from the group consisting of halogen,
cyano, nitro, tetrazolyl, guanidino, amidino, methylguanidino,
azido, --C(O)Z.sup.7, --OC(O)NH.sub.2, --OC(O)NHZ.sup.7,
--OC(O)NZ.sup.7Z.sup.8, --NHC(O)Z.sup.7, --NHC(O)NH.sub.2,
--NHC(O)NHZ.sup.7, --NHC(O)NZ.sup.7Z.sup.8, --C(O)OH,
--C(O)OZ.sup.7, --C(O)NH.sub.2, --C(O)NHZ.sup.7,
--C(O)NZ.sup.7Z.sup.8, --P(O).sub.3H.sub.2,
--P(O).sub.3(Z.sup.7).sub.2, --S(O).sub.3H, --S(O)Z.sup.7,
--S(O).sub.2 Z.sup.7, --S(O).sub.3Z.sup.7, --Z.sup.7, --OZ.sup.7,
--OH, --NH.sub.2, --NHZ.sup.7, --NZ.sup.7Z.sup.8,
--C(.dbd.NH)NH.sub.2, --C(.dbd.NOH)NH.sub.2, --N-morpholino,
(C.sub.2 C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl, (C.sub.1
C.sub.6)haloalkyl, (C.sub.2 C.sub.6)haloalkenyl, (C.sub.2
C.sub.6)haloalkynyl, (C.sub.1 C.sub.6)haloalkoxy,
--(CZ.sup.9Z.sup.10).sub.rNH.sub.2,
--(CZ.sup.9Z.sup.10).sub.rNHZ.sup.3,
--(CZ.sup.9Z.sup.10).sub.rNZ.sup.7Z.sup.8--X.sup.6(CZ.sup.9Z.sup.10).sub.-
r--(C.sub.3 C.sub.8)cycloalkyl,
--X.sup.6(CZ.sup.9Z.sup.10).sub.r--(C.sub.5 C.sub.8)cycloalkenyl,
--X.sup.6 (CZ.sup.9Z.sup.10).sub.r-aryl, and
--X.sup.6(CZ.sup.9Z.sup.10).sub.r-heterocycle; r is 1, 2, 3, or 4;
X.sup.6 is O, S, NH, --C(O)--, --C(O)NH--, --C(O)O--, --S(O)--,
--S(O).sub.2--, or --S(O).sub.3--; Z.sup.7 and Z.sup.8 are
independently selected from the group consisting of alkyl of 1 to
12 carbon atoms, alkenyl of 2 to 12 carbon atoms, alkynyl of 2 to
12 carbon atoms, cycloalkyl of 3 to 8 carbon atoms, cycloalkenyl of
5 to 8 carbon atoms, aryl of 6 to 14 carbon atoms, heterocycle of 5
to 14 ring atoms, aralkyl of 7 to 15 carbon atoms, and
heteroaralkyl of 5 to 14 ring atoms, or Z.sup.7 and Z.sup.8
together may optionally form a heterocycle; and Z.sup.9 and
Z.sup.10 are independently selected from the group consisting of
hydrogen, fluorine, alkyl of 1 to 12 carbon atoms, aryl of 6 to 14
carbon atoms, heteroaryl of about 5 to 14 ring atoms, aralkyl of 7
to 15 carbon atoms, and heteroaralkyl of 5 to 14 ring atoms, or
Z.sup.9 and Z.sup.10 are selected together to form a carbocycle, or
two Z.sup.9 groups on adjacent carbon atoms are selected together
to form a carbocycle; or (ii) any two Y.sup.2 or Y.sup.3 groups
attached to adjacent carbon atoms may be selected together to be
--O[C(Z.sup.9)(Z.sup.10)].sub.rO-- or
--O[C(Z.sup.9)(Z.sup.10)].sub.r+1--; or (iii) any two Y.sup.2 or
Y.sup.3 groups attached to the same or adjacent carbon atoms may be
selected together to form a carbocycle or heterocycle; and wherein
any of the above-mentioned substituents comprising a CH.sub.3
(methyl), CH.sub.2 (methylene), or CH (methine) group which is not
attached to a halogen, SO or SO.sub.2 group or to a N, O or S atom
optionally bears on said group a substituent selected from hydroxy,
halogen, (C.sub.1 C.sub.4)alkyl, (C.sub.1 C.sub.4)alkoxy and
--N[(C.sub.1 C.sub.4)alkyl][(C.sub.1 C.sub.4)alkyl]; or an N-oxide,
pharmaceutically acceptable prodrug, pharmaceutically active
metabolite, pharmaceutically acceptable salt, or pharmaceutically
acceptable solvate thereof.
In one embodiment, X.sup.2 is CH or CR.sup.1c; and X.sup.3 is CH,
or CR.sup.1d.
In another embodiment, X.sup.4 is O. In a particular aspect of this
embodiment, X.sup.2 is CR.sup.1c and X.sup.3 is CR.sup.1d.
In another embodiment, X.sup.1 is O. In a particular aspect of this
embodiment, X.sup.4 is O; X.sup.2 is CR.sup.1c; X.sup.3 is
CR.sup.1d and X.sup.5 is CH.
In another embodiment, X.sup.1 is CH.sub.2. In a particular aspect
of this embodiment, X.sup.4 is O; X.sup.2 is CR.sup.1c; and X.sup.3
is CR.sup.1d.
In another embodiment X.sup.5 is CH. In a particular aspect of this
embodiment, X.sup.1 is CH.sub.2. In another particular aspect of
this embodiment, X.sup.1 is CH.sup.2, X.sup.4 is O; X.sup.2 is
CR.sup.1c; and X.sup.3 is CR.sup.1d. In another particular aspect
of this embodiment, X.sup.1 is CH.sub.2, X.sup.4 is O; X.sup.2 is
CR.sup.1c; X.sup.3 is CR.sup.1d and each of R.sup.1a, R.sup.1b,
R.sup.1c, and R.sup.1d is independently H, F, or Cl. In another
particular aspect of this embodiment, X.sup.1 is CH.sub.2, X.sup.4
is O; X.sup.2 is CR.sup.1c; X.sup.3 is CR.sup.1d, each of R.sup.1a,
R.sup.1b, R.sup.1c, and R.sup.1d is independently H, F, or Cl, and
each of R.sup.4a, R.sup.4b, and R.sup.4c is independently selected
from the group consisting of H or F. In another particular aspect
of this embodiment, X.sup.1 is CH.sub.2, X.sup.4 is O; X.sup.2 is
CR.sup.1c; X.sup.3 is CR.sup.1d, each of R.sup.1a, R.sup.1b,
R.sup.1c, and R.sup.1d is independently H, F, or Cl, each of
R.sup.4a, R.sup.4b, and R.sup.4c is independently selected from the
group consisting of H or F, and R.sup.3 is either (a) a (C.sub.1
C.sub.8)alkyl, optionally substituted with 1 3 independently
selected Y.sup.2 groups; or (b) a heterocycle, optionally
substituted with 1 3 independently selected Y.sup.2 groups. In
another particular aspect of this embodiment, X.sup.1 is CH.sub.2,
X.sup.4 is O; X.sup.2 is CR.sup.1c; X.sup.3 is CR.sup.1d, each of
R.sup.1a, R.sup.1b, R.sup.1c, and R.sup.1d is independently H, F,
or Cl, each of R.sup.4a, R.sup.4b, and R.sup.4c is independently
selected from the group consisting of H or F, and R.sup.5 is either
(a) --CONR.sup.6aR.sup.6b or
--(CZ.sup.3Z.sup.4).sub.t-heterocycle.
In another embodiment, R.sup.3 is a (C.sub.1 C.sub.8)alkyl,
optionally substituted with 1 3 independently selected Y.sup.2
groups.
In another embodiment, R.sup.3 is a heterocycle, optionally
substituted with 1 3 independently selected Y.sup.2 groups.
In another embodiment, R.sup.5 is --CONR.sup.6aR.sup.6b.
In another embodiment, R.sup.5 is a
--(CZ.sup.3Z.sup.4).sub.t-heterocycle.
In another embodiment, X.sup.5 is CH; X.sup.1 is CH.sub.2; X.sup.4
is O; X.sup.2 is CR.sup.1c; X.sup.3 is CR.sup.1d; each of R.sup.1a,
R.sup.1b, R.sup.1c, and R.sup.1d is independently H, F, or Cl; and
each of R.sup.4a, R.sup.4b, and R.sup.4c is independently selected
from the group consisting of H or F; and R.sup.5 is
--CONR.sup.6aR.sup.6b.
In another embodiment are compounds having the structure of Formula
(I), wherein X.sup.5 is CH; X.sup.1 is CH.sub.2; X.sup.4 is O;
X.sup.2 is CR.sup.1c; X.sup.3 is CR.sup.1d; each of R.sup.1a,
R.sup.1b, R.sup.1c, and R.sup.1d is independently H, F, or Cl; and
each of R.sup.4a, R.sup.4b, and R.sup.4c is independently selected
from the group consisting of H or F; and R.sup.5 is
--(CZ.sup.3Z.sup.4).sub.t-heterocycle.
In another embodiment, X.sup.3 is CR.sup.1d. In a particular aspect
of this embodiment, X.sup.2 is CR.sup.1c. In another particular
aspect of this embodiment, X.sup.2 is CR.sup.1c and X.sup.5 is
CR.sup.4c. In another particular aspect of this embodiment, X.sup.2
is CR.sup.1c, X.sup.5 is CR.sup.4c, and X.sup.1 is
CR.sup.2aR.sup.2b. In another particular aspect of this embodiment,
X.sup.2 is CR.sup.1c, X.sup.5 is CR.sup.4c, X.sup.1 is
CR.sup.2aR.sup.2b, and X.sup.4 is O. In another particular aspect
of this embodiment, X.sup.2 is CR.sup.1c, X.sup.5 is CR.sup.4c,
X.sup.1 is CR.sup.2aR.sup.2b, X.sup.4 is O, and R.sup.5 is
--C(O)NR.sup.6aR.sup.6b or --(CZ.sup.3Z.sup.4).sub.t-heterocycle.
In another particular aspect of this embodiment, X.sup.2 is
CR.sup.1c, X.sup.5 is CR.sup.4c, X.sup.1 is CR.sup.2aR.sup.2b,
X.sup.4 is O, R.sup.5 is --C(O)NR.sup.6aR.sup.6b or
--(CZ.sup.3Z.sup.4).sub.t-heterocycle, and R.sup.3 is (C.sub.1
C.sub.6)alkyl or --(CZ.sup.1Z.sup.2).sub.sheterocycle. In another
particular aspect of this embodiment, X.sup.2 is CR.sup.1c, X.sup.5
is CR.sup.4c, X.sup.1 is CR.sup.2aR.sup.2b, X.sup.4 is O, R.sup.5
is --C(O)NR.sup.6aR.sup.6b or
--(CZ.sup.3Z.sup.4).sub.t-heterocycle, and R.sup.3 is heteroaryl.
In another particular aspect of this embodiment, X.sup.2 is
CR.sup.1c, X.sup.5 is CR.sup.4c, X.sup.1 is CR.sup.2aR.sup.2b,
X.sup.4 is O, R.sup.5 is --C(O)NR.sup.6aR.sup.6b or heteroaryl, and
R.sup.3 is heteroaryl. In another particular aspect of this
embodiment, X.sup.2 is CR.sup.1c, X.sup.5 is CR.sup.4c, X.sup.1 is
CR.sup.2aR.sup.2b, X.sup.4 is O, R.sup.5 is --C(O)N.sup.6aR.sup.6b,
wherein R.sup.6a and R.sup.6b taken with the nitrogen atom form a
heterocycle or R.sup.5 is imidazolye, either optionally substituted
with Y.sup.3, and R.sup.3 is heteroaryl. In another particular
aspect of this embodiment, X.sup.2 is CR.sup.1c, X.sup.5 is
CR.sup.4c, X.sup.1 is CR.sup.2aR.sup.2b, X.sup.4 is O, R.sup.5 is
--C(O)NR.sup.6aR.sup.6b, wherein R.sup.6a and R.sup.6b taken with
the nitrogen atom form a heterocycle or R.sup.5 is imidazolye,
either optionally substituted with Y.sup.3, R.sup.3 is heteroaryl,
and R.sup.1a, R.sup.1b, R.sup.1c, R.sup.1d, R.sup.2a, R.sup.2b,
R.sup.4a, R.sup.4b and R.sup.4c are each independently hydrogen or
halogen. In another particular aspect of this embodiment, X.sup.2
is CR.sup.1c, X.sup.5 is CR.sup.4c, X.sup.1 is CR.sup.2aR.sup.2b,
X.sup.4 is O, R.sup.5 is --C(O)NR.sup.6aR.sup.6b, wherein R.sup.6a
and R.sup.6b taken with the nitrogen atom form a heterocycle or
R.sup.5 is imidazolye, either optionally substituted with Y.sup.3,
R.sup.3 is heteroaryl, and R.sup.1a, R.sup.1b, R.sup.1c, R.sup.1d,
R.sup.2a, R.sup.2b, R.sup.4a, R.sup.4b and R.sup.4c are
hydrogen.
In another embodiment, X.sup.2 is CR.sup.1c, X.sup.3 is CR.sup.1d
and X.sup.5 is CR.sup.4c. In a particular aspect of this
embodiment, X.sup.4 is O. In another particular aspect of this
embodiment, X.sup.4 is O and X.sup.1 is CR.sup.2aR.sup.2b. In
another particular aspect of this embodiment, X.sup.4 is O, X.sup.1
is CR.sup.2aR.sup.2b, and R.sup.1a, R.sup.1b, R.sup.1c, R.sup.1d,
R.sup.2a, R.sup.2b, R.sup.4a, R.sup.4b and R.sup.4c are each
independently hydrogen or halogen. In another particular aspect of
this embodiment, X.sup.4 is O, X.sup.1 is CR.sup.2aR.sup.2b,
R.sup.1a, R.sup.1b, R.sup.1c, R.sup.1d, R.sup.2a, R.sup.2b,
R.sup.4a, R.sup.4b and R.sup.4c are each independently hydrogen or
halogen, and R.sup.5 is --C(O)NR.sup.6aR.sup.6b or
--(CZ.sup.3Z.sup.4).sub.t-heterocycle. In another particular aspect
of this embodiment, X.sup.4 is O, X.sup.1 is CR.sup.2aR.sup.2b,
R.sup.1a, R.sup.1b, R.sup.1c, R.sup.1d, R.sup.2a, R.sup.2b,
R.sup.4a, R.sup.4b and R.sup.4c are each independently hydrogen or
halogen, R.sup.5 is --C(O)NR.sup.6aR.sup.6b or
--(CZ.sup.3Z.sup.4).sub.t-heterocycle, and R.sup.3 is (C.sub.1
C.sub.6)alkyl or --(CZ.sup.1Z.sup.2).sub.s-heterocycle.
In another embodiment, the invention provides a compound of Formula
(II):
##STR00003## wherein (a) X.sup.1 is O or CR.sup.2aR.sup.2b; (b)
each of R.sup.1a, R.sup.1b, R.sup.1c, and R.sup.1d is independently
selected from the group consisting of hydrogen, halogen, (C.sub.1
C.sub.6)alkoxy, (C.sub.1 C.sub.6)alkyl, (C.sub.1
C.sub.6)fluoroalkoxy, and (C.sub.1 C.sub.6)fluoroalkyl; (c) each of
R.sup.2a and R.sup.2b is independently selected from H, halogen, or
a moiety, optionally substituted with 1 to 3 independently selected
Y.sup.1 groups, selected from the group consisting of (C.sub.1
C.sub.6)alkoxy, (C.sub.1 C.sub.6)alkylamine and (C.sub.1
C.sub.6)alkyl, wherein any number of the hydrogen atoms on the
(C.sub.1 C.sub.6)alkoxy and (C.sub.1 C.sub.6)alkyl groups may be
optionally replaced with F; or R.sup.2a and R.sup.2b together can
be oxo or a moiety, optionally substituted with 1 to 3
independently selected Y.sup.1 groups, selected from the group
consisting of (C.sub.3 C.sub.6)cycloalkyl, 3 6 membered
heterocycloalkyl and .dbd.CH--(C.sub.1 to C.sub.5)alkyl; (d)
R.sup.3 is H or a moiety, optionally substituted with 1 3
independently selected Y.sup.2 groups, selected from the group
consisting of --(CZ.sup.1Z.sup.2).sub.sCN,
--(CZ.sup.1Z.sup.2).sub.s--(C.sub.3 C.sub.8)cycloalkyl,
--(CZ.sup.1Z.sup.2).sub.s--(C.sub.5 C.sub.8)cycloalkenyl, (C.sub.2
C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl,
--(CZ.sup.1Z.sup.2).sub.s-aryl,
--(CZ.sup.1Z.sup.2).sub.s-heterocycle, and (C.sub.1 C.sub.8)alkyl,
where s is 0, 1, 2, or 3, and wherein when s is 2 or 3, the
CZ.sup.1Z.sup.2 units may be the same or different; (e) each of
R.sup.4a, R.sup.4b, and R.sup.4c is independently selected from the
group consisting of H, F, Cl, CF.sub.3, CH.sub.3, OCH.sub.3, and
OCF.sub.3; (f) R.sup.5 is selected from the group consisting of
hydrogen, nitro, halogen, azido, --NR.sup.6aR.sup.6b,
--NR.sup.6aSO.sub.2R.sup.6b, --NR.sup.6aC(O)R.sup.6b,
OC(O)R.sup.6b, --NR.sup.6aC(O)OR.sup.6b, --OC(O)NR.sup.6aR.sup.6b,
--OR.sup.6a, --SR.sup.6a, --S(O)R.sup.6a, --SO.sub.2R.sup.6a,
--SO.sub.3R.sup.6a, --SO.sub.2NR.sup.6aR.sup.6b, --COR.sup.6a,
--CO.sub.2R.sup.6a, --CONR.sup.6aR.sup.6b, --(C.sub.1
C.sub.4)fluoroalkyl, --(C.sub.1 C.sub.4)fluoroalkoxy,
--(CZ.sup.3Z.sup.4).sub.tCN, and a moiety selected from the group
consisting of --(CZ.sup.3Z.sup.4).sub.t-aryl,
--(CZ.sup.3Z.sup.4).sub.t-heterocycle, (C.sub.2 C.sub.6)alkynyl,
--(CZ.sup.3Z.sup.4).sub.t--(C.sub.3 C.sub.6)cycloalkyl,
--(CZ.sup.3Z.sup.4).sub.t--(C.sub.5 C.sub.6)cycloalkenyl, (C.sub.2
C.sub.6)alkenyl, and (C.sub.1 C.sub.6)alkyl, which is optionally
substituted with 1 to 3 independently selected Y.sup.2 groups,
where t is 0, 1, 2, or 3, and wherein when t is 2 or 3, the
CZ.sup.3Z.sup.4 units may be the same or different; (g) each
R.sup.6a and R.sup.6b is independently selected from the group
consisting of hydrogen and a moiety selected from the group
consisting of --(CZ.sup.5Z.sup.6).sub.u--(C.sub.3
C.sub.6)cycloalkyl, --(CZ.sup.5Z.sup.6).sub.u--(C.sub.1
C.sub.5)cycloalkenyl, --(CZ.sup.5Z.sup.6).sub.u-aryl,
--(CZ.sup.5Z.sup.6).sub.u-h eterocycle, (C.sub.2 C.sub.6)alkenyl,
and (C.sub.1 C.sub.6)alkyl, which is optionally substituted with 1
to 3 independently selected Y.sup.3 groups, where u is 0, 1, 2, or
3, and wherein when u is 2 or 3, the CZ.sup.5Z.sup.6 units may be
the same or different, or R.sup.6a and R.sup.6b taken together can
with adjacent atoms form a heterocycle; (h) each Z.sup.1, Z.sup.2,
Z.sup.3, Z.sup.4, Z.sup.5, and Z.sup.6 is independently selected
from the group consisting of H, F, and (C.sub.1 C.sub.6)alkyl, or
each Z.sup.1 and Z.sup.2, Z.sup.3 and Z.sup.4, or Z.sup.5 and
Z.sup.6 are selected together to form a carbocycle, or two Z.sup.1,
Z.sup.3 or Z.sup.3 groups on adjacent carbon atoms are selected
together to optionally form a carbocycle; and (i) each Y.sup.1 is
independently selected from the group consisting of halogen, cyano,
nitro, azido, --OH, --NH.sub.2, (C.sub.1 C.sub.6)alkoxy, (C.sub.1
C.sub.6)alkylamino, (C.sub.1 C.sub.6)dialkylamino, (C.sub.1
C.sub.6)alkyl, (C.sub.2 C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl,
(C.sub.1 C.sub.6)haloalkyl, (C.sub.1 C.sub.6)haloalkoxy, --(C.sub.3
C.sub.6)cycloalkyl; (j) each Y.sup.2 and Y.sup.3 is independently
selected and (i) is selected from the group consisting of halogen,
cyano, nitro, tetrazolyl, guanidino, amidino, methylguanidino,
azido, --C(O)Z.sup.7, --OC(O)NH.sub.2, --OC(O)NHZ.sup.7,
--OC(O)NZ.sup.7Z.sup.8, --NHC(O)Z.sup.7, --NHC(O)NH.sub.2,
--NHC(O)NHZ.sup.7, --NHC(O)NZ.sup.7Z.sup.8, --C(O)OH,
--C(O)OZ.sup.7, --C(O)NH.sub.2, --C(O)NHZ.sup.7,
--C(O)NZ.sup.7Z.sup.8, --P(O).sub.3H.sub.2,
--P(O).sub.3(Z.sup.7).sub.2, --S(O).sub.3H, --S(O)Z.sup.7,
--S(O).sub.2Z.sup.7, --S(O).sub.3Z.sup.7, --Z.sup.7, --OZ.sup.7,
--OH, --NH.sub.2, --NHZ.sup.7, --NZ.sup.7Z.sup.8,
--C(.dbd.NH)NH.sub.2, --C(.dbd.NOH)NH.sub.2, --N-morpholino,
(C.sub.2 C.sub.8)alkenyl, (C.sub.2 C.sub.6)alkynyl, (C.sub.1
C.sub.6)haloalkyl, (C.sub.2 C.sub.6)haloalkenyl, (C.sub.2
C.sub.6)haloalkynyl, (C.sub.1 C.sub.6)haloalkoxy,
--(CZ.sup.9Z.sup.10).sub.rNH.sub.2,
--(CZ.sup.9Z.sup.10).sub.rNHZ.sup.3,
--(CZ.sup.9Z.sup.10).sub.rNZ.sup.7Z.sup.8--X.sup.6(CZ.sup.9Z.sup.10).sub.-
r--(C.sub.3 C.sub.8) cycloalkyl,
--X.sup.6(CZ.sup.9Z.sup.10).sub.r--(C.sub.5 C.sub.8)cycloalkenyl,
--X.sup.6(CZ.sup.9Z.sup.10).sub.r-aryl, and
--X.sup.6(CZ.sup.9Z.sup.10).sub.r-heterocycle; r is 1, 2, 3, or 4;
X.sup.6 is O, S, NH, --C(O)--, --C(O)NH--, --C(O)O--, --S(O)--,
--S(O).sub.2--, or --S(O).sub.3--; Z.sup.7 and Z.sup.8 are
independently selected from the group consisting of alkyl of 1 to
12 carbon atoms, alkenyl of 2 to 12 carbon atoms, alkynyl of 2 to
12 carbon atoms, cycloalkyl of 3 to 8 carbon atoms, cycloalkenyl of
5 to 8 carbon atoms, aryl of 6 to 14 carbon atoms, heterocycle of 5
to 14 ring atoms, aralkyl of 7 to 15 carbon atoms, and
heteroaralkyl of 5 to 14 ring atoms, or Z.sup.7 and Z.sup.8
together may optionally form a heterocycle; and Z.sup.9 and
Z.sup.10 are independently selected from the group consisting of
hydrogen, fluorine, alkyl of 1 to 12 carbon atoms, aryl of 6 to 14
carbon atoms, heteroaryl of about 5 to 14 ring atoms, aralkyl of 7
to 15 carbon atoms, and heteroaralkyl of 5 to 14 ring atoms, or
Z.sup.9 and Z.sup.10 are selected together to form a carbocycle, or
two Z.sup.9 groups on adjacent carbon atoms are selected together
to form a carbocycle; or (ii) any two Y.sup.2 or Y.sup.3 groups
attached to adjacent carbon atoms may be selected together to be
--O[C(Z.sup.9)(Z.sup.10)].sub.rO-- or
--O[C(Z.sup.9)(Z.sup.10)].sub.r+1--; or (iii) any two Y.sup.2 or
Y.sup.3 groups attached to the same or adjacent carbon atoms may be
selected together to form a carbocycle or heterocycle; and wherein
any of the above-mentioned substituents comprising a CH.sub.3
(methyl), CH.sub.2 (methylene), or CH (methine) group which is not
attached to a halogen, SO or SO.sub.2 group or to a N, O or S atom
optionally bears on said group a substituent selected from hydroxy,
halogen, (C.sub.1 C.sub.4)alkyl, (C.sub.1 C.sub.4)alkoxy and
--N[(C.sub.1 C.sub.4)alkyl][(C.sub.1 C.sub.4)alkyl]; or a
pharmaceutically acceptable salt or solvate thereof.
Preferred aspects of this embodiment include those described above
for Formula I, to the extent not inconsistent with Formula II.
In another embodiment, the invention provides a compound selected
from the group consisting of:
N-(4,6-Dimethyl-pyridin-2-yl)-2-{3-fluoro-4-[2-(1-methyl-1H-imidazol-2-yl-
)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide, 2-{4-[2-((3R,
4R)-3,4-Dihydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-p-
henyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide,
2-{4-[2-((R)-3-Dimethylamino-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-
-7-yloxy]-phenyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide,
7-{4-[(4,6-Dimethyl-pyridin-2-ylcarbamoyl)-methyl]-phenoxy}-thieno[3,2-b]-
pyridine-2-carboxylic acid dimethylamide,
N-(4,6-Dimethyl-pyridin-2-yl)-2-{4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[-
3,2-b]pyridin-7-yloxy]-phenyl}-acetamide,
N-(5-Chloro-pyridin-2-yl)-2-{4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2--
b]pyridin-7-yloxy]-phenyl}-acetamide,
N-(4,6-Dimethyl-pyridin-2-yl)-2-{4-[2-((R)-3-hydroxy-pyrrolidine-1-carbon-
yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide,
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylo-
xy]-phenyl}-N-isoquinolin-3-yl-acetamide,
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylo-
xy]-phenyl}-N-phenyl-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(4-
,6-dimethyl-pyridin-2-yl)-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl-N-(6--
methyl-pyridin-2-yl)-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5-
-trifluoromethyl-pyridin-2-yl)-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5-
-chloro-pyridin-2-yl)-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5-
-bromo-pyridin-2-yl)-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-is-
oquinolin-3-yl-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phe-
nyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide,
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phe-
nyl}-N-(5-methyl-1H-pyrazol-3-yl)-acetamide, and Butyl-carbamic
acid
4-[2-(azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl
ester, or a pharmaceutically acceptable prodrug, pharmaceutically
active metabolite, pharmaceutically acceptable solvate or
pharmaceutically acceptable salt thereof.
In another embodiment, the invention provides a compound selected
from the group consisting of:
##STR00004## ##STR00005## ##STR00006## ##STR00007## or a
pharmaceutically acceptable prodrug, pharmaceutically active
metabolite, pharmaceutically acceptable solvate or pharmaceutically
acceptable salt thereof.
In another aspect of the present invention are methods for
producing a compound having the structure of Formula (I), wherein
X.sup.2 is CR.sup.1c, comprising: (a) reacting a carboxylic acid
having the structure
##STR00008## with a chlorinating agent; and (b) reacting the
corresponding product with H.sub.2N--R.sup.3. In a further
embodiment of this method, the chlorinating agent is selected from
the group consisting of thionyl chloride, oxalyl chloride, and
chlorine.
In a further embodiment are methods for producing the the
carboxylic acid having the structure:
##STR00009## comprising (a) reacting a compound having the
formula
##STR00010## with a compound having the formula
##STR00011## in the presence of a base.
In another embodiment are methods for producing a compound having
the structure of Formula (I), wherein X.sup.2 is O ***?? X4 is O?
*** comprising (a) reacting
##STR00012## with a carbonyl electrophile; and (b) reacting the
corresponding product with H.sub.2N--R.sup.3. In a further
embodiment of this method the carbonyl electrophile is
phosgene.
This invention also relates to a pharmaceutical composition for the
treatment of abnormal cell growth in a mammal, including a human,
comprising an amount of a compound of the formula 1, as defined
above, or a pharmaceutically acceptable salt, solvate or prodrug
thereof, that is effective in treating abnormal cell growth, and a
pharmaceutically acceptable carrier. In one embodiment of said
composition, said abnormal cell growth is cancer, including, but
not limited to, lung cancer, bone cancer, pancreatic cancer, skin
cancer, cancer of the head or neck, cutaneous or intraocular
melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of
the anal region, stomach cancer, colon cancer, breast cancer,
uterine cancer, carcinoma of the fallopian tubes, carcinoma of the
endometrium, carcinoma of the cervix, carcinoma of the vagina,
carcinoma of the vulva, Hodgkin's Disease, cancer of the esophagus,
cancer of the small intestine, cancer of the endocrine system,
cancer of the thyroid gland, cancer of the parathyroid gland,
cancer of the adrenal gland, sarcoma of soft tissue, cancer of the
urethra, cancer of the penis, prostate cancer, chronic or acute
leukemia, lymphocytic lymphomas, cancer of the bladder, cancer of
the kidney or ureter, renal cell carcinoma, carcinoma of the renal
pelvis, neoplasms of the central nervous system (CNS), primary CNS
lymphoma, spinal axis tumors, brain stem glioma, pituitary adenoma,
or a combination of one or more of the foregoing cancers. In
another embodiment of said pharmaceutical composition, said
abnormal cell growth is a benign proliferative disease, including,
but not limited to, psoriasis, benign prostatic hypertrophy or
restinosis.
The invention also relates to a pharmaceutical composition for the
treatment of abnormal cell growth in a mammal, including a human,
which comprises an amount of a compound of formula 1, as defined
above, or a pharmaceutically acceptable salt, solvate or prodrug
thereof, that is effective in treating abnormal cell growth in
combination with a pharmaceutically acceptable carrier and an
anti-tumor agent selected from the group consisting of mitotic
inhibitors, alkylating agents, anti-metabolites, intercalating
antibiotics, growth factor inhibitors, cell cycle inhibitors,
enzymes, topoisomerase inhibitors, biological response modifiers,
anti-hormones, and anti-androgens.
This invention also relates to a method for the treatment of
abnormal cell growth in a mammal, including a human, comprising
administering to said mammal an amount of a compound of the formula
1, as defined above, or a pharmaceutically acceptable salt or
solvate thereof, that is effective in treating abnormal cell
growth. In one embodiment of this method, the abnormal cell growth
is cancer, including, but not limited to, lung cancer, bone cancer,
pancreatic cancer, skin cancer, cancer of the head or neck,
cutaneous or intraocular melanoma, uterine cancer, ovarian cancer,
rectal cancer, cancer of the anal region, stomach cancer, colon
cancer, breast cancer, uterine cancer, carcinoma of the fallopian
tubes, carcinoma of the endometrium, carcinoma of the cervix,
carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease,
cancer of the esophagus, cancer of the small intestine, cancer of
the endocrine system, cancer of the thyroid gland, cancer of the
parathyroid gland, cancer of the adrenal gland, sarcoma of soft
tissue, cancer of the urethra, cancer of the penis, prostate
cancer, chronic or acute leukemia, lymphocytic lymphomas, cancer of
the bladder, cancer of the kidney or ureter, renal cell carcinoma,
carcinoma of the renal pelvis, neoplasms of the central nervous
system (CNS), primary CNS lymphoma, spinal axis tumors, brain stem
glioma, pituitary adenoma, or a combination of one or more of the
foregoing cancers. In another embodiment of said method, said
abnormal cell growth is a benign proliferative disease, including,
but not limited to, psoriasis, benign prostatic hypertrophy or
restinosis.
This invention also relates to a method for the treatment of a
disorder associated with angiogenesis in a mammal, including a
human, comprising administering to said mammal an amount of a
compound of the formula 1, as defined above, or a pharmaceutically
acceptable salt, solvate or prodrug thereof, that is effective in
treating said disorder. Such disorders include cancerous tumors
such as melanoma; ocular disorders such as age-related macular
degeneration, presumed ocular histoplasmosis syndrome, and retinal
neovascularization from proliferative diabetic retinopathy;
rheumatoid arthritis; bone loss disorders such as osteoporosis,
Paget's disease, humoral hypercalcemia of malignancy, hypercalcemia
from tumors metastatic to bone, and osteoporosis induced by
glucocorticoid treatment; coronary restenosis; and certain
microbial infections including those associated with microbial
pathogens selected from adenovirus, hantaviruses, Borrelia
burgdorferi, Yersinia spp., Bordetella pertussis, and group A
Streptococcus.
This invention also relates to a method of (and to a pharmaceutical
composition for) treating abnormal cell growth in a mammal which
comprise an amount of a compound of formula 1, or a
pharmaceutically acceptable salt, solvate or prodrug thereof, in
combination with an amount of one or more substances selected from
anti-tumor agents, anti-angiogenesis agents, signal transduction
inhibitors, and antiproliferative agents, which amounts are
together effective in treating said abnormal cell growth. Such
substances include those disclosed in PCT publication Nos. WO
00/38715, WO 00/38716, WO 00/38717, WO 00/38718, WO 00/38719, WO
00/38730, WO 00/38665, WO 00/37107 and WO 00/38786, all published
on Jul. 6, 2000, the disclosures of which are incorporated herein
by reference in their entireties for all purposes.
Anti-tumor agents can be used in conjunction with a compound of
formula 1 in the methods and pharmaceutical compositions described
herein. Examples of anti-tumor agents include mitotic inhibitors,
for example vinca alkaloid derivatives such as vinblastine
vinorelbine, vindescine and vincristine; colchines allochochine,
halichondrine, N-benzoyltrimethyl-methyl ether colchicinic acid,
dolastatin 10, maystansine, rhizoxine, taxanes such as taxol
(paclitaxel), docetaxel (Taxotere),
2'-N-[3-(dimethylamino)propyl]glutaramate (taxol derivative),
thiocholchicine, trityl cysteine, teniposide, methotrexate,
azathioprine, fluorouricil, cytocine arabinoside,
2'2'-difluorodeoxycytidine (gemcitabine), adriamycin and mitamycin.
Alkylating agents, for example cis-platin, carboplatin oxiplatin,
iproplatin, Ethyl ester of N-acetyl-DL-sarcosyl-L-leucine (Asaley
or Asalex), 1,4-cyclohexadiene-1,4-dicarbamic acid,
2,5-bis(1-azirdinyl)-3,6-dioxo-, diethyl ester (diaziquone),
1,4-bis(methanesulfonyloxy)butane (bisulfan or leucosulfan)
chlorozotocin, clomesone, cyanomorpholinodoxorubicin, cyclodisone,
dianhydroglactitol, fluorodopan, hepsulfam, mitomycin C,
hycantheonemitomycin C, mitozolamide,
1-(2-chloroethyl)-4-(3-chloropropyl)-piperazine dihydrochloride,
piperazinedione, pipobroman, porfiromycin, spirohydantoin mustard,
teroxirone, tetraplatin, thiotepa, triethylenemelamine, uracil
nitrogen mustard, bis(3-mesyloxypropyl)amine hydrochloride,
mitomycin, nitrosoureas agents such as
cyclohexyl-chloroethylnitrosourea,
methylcyclohexyl-chloroethylnitrosourea
1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidyl)-1-nitroso-urea,
bis(2-chloroethyl)nitrosourea, procarbazine, dacarbazine, nitrogen
mustard-related compounds such as mechloroethamine,
cyclophosphamide, ifosamide, melphalan, chlorambucil, estramustine
sodium phosphate, strptozoin, and temozolamide. DNA
anti-metabolites, for example 5-fluorouracil, cytosine arabinoside,
hydroxyurea,
2-[(3hydroxy-2-pyrinodinyl)methylene]-hydrazinecarbothioamide,
deoxyfluorouridine, 5-hydroxy-2-formylpyridine thiosemicarbazone,
alpha-2'-deoxy-6-thioguanosine, aphidicolin glycinate,
5-azadeoxycytidine, beta-thioguanine deoxyriboside, cyclocytidine,
guanazole, inosine glycodialdehyde, macbecin II, pyrazolimidazole,
cladribine, pentostatin, thioguanine, mercaptopurine, bleomycin,
2-chlorodeoxyadenosine, inhibitors of thymidylate synthase such as
raltitrexed and pemetrexed disodium, clofarabine, floxuridine and
fludarabine. DNA/RNA antimetabolites, for example, L-alanosine,
5-azacytidine, acivicin, aminopterin and derivatives thereof such
as
N-[2-chloro-5-[[(2,4-diamino-5-methyl-6-quinazolinyl)methyl]amino]benzoyl-
]-L-aspartic acid,
N-[4-[[(2,4-diamino-5-ethyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspart-
ic acid,
N-[2-chloro-4-[[(2,4-diaminopteridinyl)methyl]amino]benzoyl]-L-as-
partic acid, soluble Baker's antifol, dichloroallyl lawsone,
brequinar, ftoraf, dihydro-5-azacytidine, methotrexate,
N-(phosphonoacetyl)-L-aspartic acid tetrasodium salt, pyrazofuran,
trimetrexate, plicamycin, actinomycin D, cryptophycin, and analogs
such as cryptophycin-52 or, for example, one of the preferred
anti-metabolites disclosed in European Patent Application No.
239362 such as
N-(5-[N-(3,4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl)-N-methylamino]--
2-thenoyl)-L-glutamic acid; growth factor inhibitors; cell cycle
inhibitors; intercalating antibiotics, for example adriamycin and
bleomycin; proteins, for example interferon; and anti-hormones, for
example anti-estrogens such as Nolvadex.TM. (tamoxifen) or, for
example anti-androgens such as Casodex.TM.
(4'-cyano-3-(4-fluorophenylsulphonyl)-2-hydroxy-2-methyl-3'-(trifluoromet-
hyl)propionanilide). Such conjoint treatment may be achieved by way
of the simultaneous, sequential or separate dosing of the
individual components of the treatment.
Anti-angiogenesis agents, such as MMP-2 (matrix-metalloprotienase
2) inhibitors, MMP-9 (matrix-metalloprotienase 9) inhibitors, and
COX-II (cyclooxygenase II) inhibitors, can be used in conjunction
with a compound of formula 1 in the methods and pharmaceutical
compositions described herein. Examples of useful COX-II inhibitors
include CELEBREX.TM. (alecoxib), valdecoxib, and rofecoxib.
Examples of useful matrix metalloproteinase inhibitors are
described in WO 96/33172 (published Oct. 24, 1996), WO 96/27583
(published Mar. 7, 1996), European Patent Application No.
97304971.1 (filed Jul. 8, 1997), European Patent Application No.
99308617.2 (filed Oct. 29, 1999), WO 98/07697 (published Feb. 26,
1998), WO 98/03516 (published Jan. 29, 1998), WO 98/34918
(published Aug. 13, 1998), WO 98/34915 (published Aug. 13, 1998),
WO 98/33768 (published Aug. 6, 1998), WO 98/30566 (published Jul.
16, 1998), European Patent Publication 606,046 (published Jul. 13,
1994), European Patent Publication 931,788 (published Jul. 28,
1999), WO 90/05719 (published May 331, 1990), WO 99/52910
(published Oct. 21, 1999), WO 99/52889 (published Oct. 21, 1999),
WO 99/29667 (published Jun. 17, 1999), PCT International
Application No. PCT/IB98/01113 (filed Jul. 21, 1998), European
Patent Application No. 99302232.1 (filed Mar. 25, 1999), Great
Britain patent application number 9912961.1 (filed Jun. 3, 1999),
U.S. Provisional Application No. 60/148,464 (filed Aug. 12, 1999),
U.S. Pat. No. 5,863,949 (issued Jan. 26, 1999), U.S. Pat. No.
5,861,510 (issued Jan. 19, 1999), and European Patent Publication
780,386 (published Jun. 25, 1997), all of which are herein
incorporated by reference in their entirety. Preferred MMP-2 and
MMP-9 inhibitors are those that have little or no activity
inhibiting MMP-1. More preferred, are those that selectively
inhibit MMP-2 and/or MMP-9 relative to the other
matrix-metalloproteinases (i.e. MMP-1, MMP-3, MMP4, MMP-5, MMP-6,
MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13).
Some specific examples of MMP inhibitors useful in combination with
the compounds of the present invention are AG-3340, RO 32-3555, RS
13-0830, and the compounds recited in the following list:
3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl-
)-amino]-propionic acid;
3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]o-
ctane-3-carboxylic acid hydroxyamide; (2R, 3R)
1-[4-(2-chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-pi-
peridine-2-carboxylic acid hydroxyamide;
4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxyl-
ic acid hydroxyamide;
3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-
-amino]-propionic acid;
4-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxyl-
ic acid hydroxyamide;
3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxyl-
ic acid hydroxyamide; (2R, 3R)
1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-pi-
peridine-2-carboxylic acid hydroxyamide;
3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-et-
hyl)-amino]-propionic acid;
3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro--
pyran-4-yl)-amino]-propionic acid;
3-exo-3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]o-
ctane-3-carboxylic acid hydroxyamide;
3-endo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]-
octane-3-carboxylic acid hydroxyamide; and
3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-furan-3-carboxyl-
ic acid hydroxyamide; and pharmaceutically acceptable salts,
solvates and prodrugs of said compounds.
Signal transduction inhibitors can be used in conjunction with a
compound of formula 1 in the methods and pharmaceutical
compositions described herein. Examples of signal transduction
inhibitors include agents that can inhibit EGFR (epidermal growth
factor receptor) responses, such as EGFR antibodies, EGF
antibodies, and molecules that are EGFR inhibitors; VEGF (vascular
endothelial growth factor) inhibitors; and erbB2 receptor
inhibitors, such as organic molecules or antibodies that bind to
the erbB2 receptor, for example, HERCEPTIN.TM. (Genentech, Inc. of
South San Francisco, Calif., USA).
EGFR inhibitors are described in, for example in WO 95/19970
(published Jul. 27, 1995), WO 98/14451 (published Apr. 9, 1998), WO
98/02434 (published Jan. 22, 1998), and U.S. Pat. No. 5,747,498
(issued May 5, 1998). EGFR-inhibiting agents include, but are not
limited to, the monoclonal antibodies C225 and anti-EGFR 22Mab
(ImClone Systems Incorporated of New York, N.Y., USA), the
compounds ZD-1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim),
MDX447 (Medarex Inc. of Annandale, N.J., USA), and OLX-103 (Merck
& Co. of Whitehouse Station, N.J., USA), VRCTC-310 (Ventech
Research) and EGF fusion toxin (Seragen Inc. of Hopkinton,
Mass.).
VEGF inhibitors, for example SU-5416 and SU-6668 (Sugen Inc. of
South San Francisco, Calif., USA), can also be combined with a
compound of formula 1. VEGF inhibitors are described in, for
example, U.S. Pat. No. 6,534,524, issued Mar. 18, 2003, U.S. Pat.
No. 6,531,491, issued Mar. 11, 2003, WO 99/24440 (published May 20,
1999), PCT International Application PCT/IB99/00797 (filed May 3,
1999), in WO 95/21613 (published Aug. 17, 1995), WO 99/61422
(published Dec. 2, 1999), U.S. Pat. No. 5,834,504 (issued Nov. 10,
1998), WO 98/50356 (published Nov. 12, 1998), U.S. Pat. No.
5,883,113 (issued Mar. 16, 1999), U.S. Pat. No. 5,886,020 (issued
Mar. 23, 1999), U.S. Pat. No. 5,792,783 (issued Aug. 11, 1998), WO
99/10349 (published Mar. 4, 1999), WO 97/32856 (published Sep. 12,
1997), WO 97/22596 (published Jun. 26, 1997), WO 98/54093
(published Dec. 3, 1998), WO 98/02438 (published Jan. 22, 1998), WO
99/16755 (published Apr. 8, 1999), and WO 98/02437 (published Jan.
22, 1998), all of which are herein incorporated by reference in
their entirety. Other examples of some specific VEGF inhibitors are
IM862 (Cytran Inc. of Kirkland, Wash., USA); anti-VEGF monoclonal
antibody of Genentech, Inc. of South San Francisco, Calif.; and
angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colo.) and
Chiron (Emeryville, Calif.).
ErbB2 receptor inhibitors, such as GW-282974 (Glaxo Wellcome plc),
and the monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc.
of The Woodlands, Tex., USA) and 2B-1 (Chiron), may be administered
in combination with a compound of formula 1. Such erbB2 inhibitors
include those described in WO 98/02434 (published Jan. 22, 1998),
WO 99/35146 (published Jul. 15, 1999), WO 99/35132 (published Jul.
15, 1999), WO 98/02437 (published Jan. 22, 1998), WO 97/13760
(published Apr. 17, 1997), WO 95/19970 (published Jul. 27, 1995),
U.S. Pat. No. 5,587,458 (issued Dec. 24, 1996), and U.S. Pat. No.
5,877,305 (issued Mar. 2, 1999), each of which is herein
incorporated by reference in its entirety. ErbB2 receptor
inhibitors useful in the present invention are also described in
U.S. Provisional Application No. 60/117,341, filed Jan. 27, 1999,
and in U.S. Provisional Application No. 60/117,346, filed Jan. 27,
1999, both of which are herein incorporated by reference in their
entirety.
Other antiproliferative agents that may be used with the compounds
of the present invention include inhibitors of the enzyme farnesyl
protein transferase and inhibitors of the receptor tyrosine kinase
PDGFr, including the compounds disclosed and claimed in the
following United States patent applications: Ser. No. 09/221,946
(filed Dec. 28, 1998); Ser. No. 09/454,058 (filed Dec. 2, 1999);
Ser. No. 09/501,163 (filed Feb. 9, 2000); Ser. No. 09/539,930
(filed Mar. 31, 2000); Ser. No. 09/202,796 (filed May 22, 1997);
Ser. No. 09/384,339 (filed Aug. 26, 1999); and Ser. No. 09/383,755
(filed Aug. 26, 1999); and the compounds disclosed and claimed in
the following United States provisional patent applications: No.
60/168,207 (filed Nov. 30, 1999); No. 60/170,119 (filed Dec. 10,
1999); No. 60/177,718 (filed Jan. 21, 2000); No. 60/168,217 (filed
Nov. 30, 1999), and No. 60/200,834 (filed May 1, 2000). Each of the
foregoing patent applications and provisional patent applications
is herein incorporated by reference in their entirety.
A compound of formula 1 may also be used with other agents useful
in treating abnormal cell growth or cancer, including, but not
limited to, agents capable of enhancing antitumor immune responses,
such as CTLA4 (cytotoxic lymphocite antigen 4) antibodies, and
other agents capable of blocking CTLA4; and anti-proliferative
agents such as other farnesyl protein transferase inhibitors, for
example the farnesyl protein transferase inhibitors described in
the references cited in the "Background" section, supra. Specific
CTLA4 antibodies that can be used in the present invention include
those described in U.S. Provisional Application No. 60/113,647
(filed Dec. 23, 1998) which is herein incorporated by reference in
its entirety.
The invention also relates to a pharmaceutical composition for the
treatment of pancreatitis or kidney disease (including
proliferative glomerulonephritis and diabetes-induced renal
disease) in a mammal which comprises a therapeutically effective
amount of a compound of formula (I), or prodrugs thereof,
pharmaceutically active metabolites, pharmaceutically acceptable
salts, or pharmaceutically acceptable solvates of said compounds
and said prodrugs, and a pharmaceutically acceptable carrier.
The invention also relates to a pharmaceutical composition for the
prevention of blastocyte implantation in a mammal which comprises a
therapeutically effective amount of a compound of formula (I), or
prodrugs thereof, pharmaceutically active metabolites,
pharmaceutically acceptable salts, or pharmaceutically acceptable
solvates of said compounds and said prodrugs, and a
pharmaceutically acceptable carrier.
The invention also relates to a pharmaceutical composition for
treating a disease related to vasculogenesis or angiogenesis in a
mammal which comprises a therapeutically effective amount of a
compound of formula (I), or prodrugs thereof, pharmaceutically
active metabolites, pharmaceutically acceptable salts, or
pharmaceutically acceptable solvates of said compounds and said
prodrugs, and a pharmaceutically acceptable carrier. In one
embodiment, said pharmaceutical composition is for treating a
disease selected from the group consisting of tumor angiogenesis,
chronic inflammatory disease such as rheumatoid arthritis,
atherosclerosis, skin diseases such as psoriasis, eczema, and
scleroderma, diabetes, diabetic retinopathy, retinopathy of
prematurity, age-related macular degeneration, hemangioma, glioma,
melanoma, Kaposi's sarcoma and ovarian, breast, lung, pancreatic,
prostate, colon and epidermoid cancer.
The invention also relates to a method of treating a
hyperproliferative disorder in a mammal which comprises
administering to said mammal a therapeutically effective amount of
the compound of formula (I), or prodrugs thereof, pharmaceutically
active metabolites, pharmaceutically acceptable salts, or
pharmaceutically acceptable solvates of said compounds and said
prodrugs in one embodiment, said method relates to the treatment of
cancer such as brain, ophthalmic, squamous cell, bladder, gastric,
pancreatic, breast, head, neck, oesophageal, prostate, colorectal,
lung, renal, kidney, ovarian, gynecological or thyroid cancer. In
another embodiment, said method relates to the treatment of a
non-cancerous hyperproliferative disorder such as benign
hyperplasia of the skin (e.g., psoriasis) or prostate (e.g.,
BPH).
The invention also relates to a method for the treatment of a
hyperproliferative disorder in a mammal which comprises
administering to said mammal a therapeutically effective amount of
a compound of formula (I), or prodrugs thereof, pharmaceutically
active metabolites, pharmaceutically acceptable salts, or
pharmaceutically acceptable solvates of said compounds and said
prodrugs, in combination with an anti-tumor agent selected from the
group consisting of mitotic inhibitors, alkylating agents,
anti-metabolites, intercalating antibiotics, growth factor
inhibitors, cell cycle inhibitors, enzymes, topoisomerase
inhibitors, biological response modifiers, anti-hormones, and
anti-androgens.
The treatment of a hyperproliferative disorder in a mammal which
comprises administering to said mammal a therapeutically effective
amount of a VEGF receptor tyrosine kinase inhibitor may lead to a
sustained increase in blood pressure. The compounds of the present
invention may be used in conjunction with an anti-hypertensive,
such as NORVASC or PROCARDIA XL, commercially available from
Pfizer, for use in the treatment of a hyperproliferative disorder
in a mammal.
This invention also relates to a pharmaceutical composition for
treating a disease related to vasculogenesis or angiogenesis in a
mammal comprising (a) therapeutically effective amount of a
compound of formula (I), or prodrugs thereof, pharmaceutically
active metabolites, pharmaceutically acceptable salts, or
pharmaceutically acceptable solvates of said compounds and said
prodrugs, (b) a therapeutically effective amount of a compound,
prodrug, metabolite, salt or solvate of an inhibitor of tumor
necrosis factor alpha, and (c) a pharmaceutically acceptable
carrier.
This invention also relates to a pharmaceutical composition for
treating a disease related to undesired angiogenesis, endothelial
cell migration or endothelial cell proliferation in a mammal
comprising (a) therapeutically effective amount of a compound of
formula (I), or prodrugs thereof, pharmaceutically active
metabolites, pharmaceutically acceptable salts, or pharmaceutically
acceptable solvates of said compounds and said prodrugs, (b) a
therapeutically effective amount of a compound, prodrug,
metabolite, salt or solvate of a NADPH oxidase inhibitor, and (c) a
pharmaceutically acceptable carrier.
This invention also relates to a pharmaceutical composition for
inhibiting abnormal cell growth in a mammal, including a human,
comprising an amount of a compound of formula (I), or prodrugs
thereof, pharmaceutically active metabolites, pharmaceutically
acceptable salts, or pharmaceutically acceptable solvates of said
compounds and said prodrugs, that is effective in inhibiting
farnesyl protein transferase, and a pharmaceutically acceptable
carrier.
This invention also relates to a pharmaceutical composition for
inhibiting abnormal cell growth in a mammal which comprises an
amount of a compound of formula (I), or prodrugs thereof,
pharmaceutically active metabolites, pharmaceutically acceptable
salts, or pharmaceutically acceptable solvates of said compounds
and said prodrugs, in combination with an amount of a
chemotherapeutic, wherein the amounts of the compound, salt,
solvate, or prodrug of formula (I), and of the chemotherapeutic are
together effective in inhibiting abnormal cell growth. Many
chemotherapeutics are presently known in the art. In one
embodiment, the chemotherapeutic is selected from the group
consisting of mitotic inhibitors, alkylating agents,
anti-metabolites, intercalating antibiotics, growth factor
inhibitors, cell cycle inhibitors, enzymes, topoisomerase
inhibitors, biological response modifiers, anti-hormones, e.g.
anti-androgens.
The compounds described herein may be used in a method for
preventing or reducing the growth of tumor cells expressing
functional VEGF-1 receptors by administering an effective amount of
a small molecule VEGF-1 receptor antagonist to inhibit autocrine
stimulation and an effective amount of a compound of Formula (I).
Active ingredients in such compositions may be present in free form
or in the form of a pharmaceutical acceptable salt and optionally
at least one pharmaceutically acceptable carrier.
The compounds described herein also may be used in combination with
a selective COX-2-inhibitor for simultaneous, separate or
sequential use. The compounds described herein may also be used in
combination with a truncated, soluble Flkl/KDR receptor to treat a
subjects having disease or disorder associated with VEGF. Active
ingredients in such compositions may be present in free form or in
the form of a pharmaceutical acceptable salt and optionally at
least one pharmaceutically acceptable carrier.
The compounds described herein also may be used in combination with
a second active ingredient which decreases the activity of, binds
to, or inhibits the epidermal growth factor (EGF). Active
ingredients in such compositions may be present in free form or in
the form of a pharmaceutical acceptable salt and optionally at
least one pharmaceutically acceptable carrier.
The compounds described herein also may be used to inhibit
VEGF-mediated angiogenesis in a tissue via several methods
including but not limited to, contacting the tissue with an
inhibitor of NADPH oxidase and an effective amount of a compound of
formula 1, by contacting the tissue with an inhibitor of reactive
oxygen species (ROS) and an effective amount of a compound of
Formula (I), or by contacting the tissue with an inhibitor of
superoxide dismutase (SOD) and an effective amount of a compound of
formula 1. Active ingredients in such compositions may be present
in free form or in the form of a pharmaceutical acceptable salt and
optionally at least one pharmaceutically acceptable carrier.
The compounds described herein may also be used in combination with
molecules which specifically bind to placenta growth factor in
order to suppress or prevent placenta growth factor-induced
pathological angiogenesis, vascular leakage (oedema), pulmonary
hypertension, tumour formation and/or inflammatory disorders.
The compounds described herein also may be used in combination with
molecules chosen from the group comprising: an antibody or any
fragment thereof which specifically binds to placenta growth
factor, a small molecule specifically binding to placenta growth
factor or to vascular endothelial growth factor receptor-1,
-vascular endothelial growth factor receptor-1 antagonists or any
fragment thereof, -a ribozyme against nucleic acids encoding
placenta growth factor or the vascular endothelial growth factor
receptor-1, and -anti-sense nucleic acids hybridizing with nucleic
acids encoding placenta growth factor or vascular endothelial
growth factor receptor-1. Active ingredients in such compositions
may be present in free form or in the form of a pharmaceutical
acceptable salt and optionally at least one pharmaceutically
acceptable carrier.
The compounds described herein may be used in a method of
inhibiting the growth of non-solid tumor cells that are stimulated
by a ligand of vascular endothelial growth factor receptor
(including but not limited to VEGFR2 kinase) in mammals, the method
comprising treating the mammals with an effective amount of a
compound of Formula (I). The compounds described herein may be used
in a method of inhibiting the growth of non-solid tumors that are
stimulated by a ligand of vascular endothelial growth factor
receptor (including but not limited to VEGFR2 kinase) in mammals,
the method comprising treating the mammals with an effective amount
of a compound of Formula (I) in combination with radiation.
The compounds described herein may also be used in combination with
G2/M agents and with therapeutic agents whose therapeutic
effectiveness is dependent, at least in part, on the presence of an
internalizing cell surface structure on the target cell. Such G2/M
agents include but are not limited to vinorelbine tartrate,
cisplatin, carboplatin, paclitaxel, doxorubicin, 5FU, docetaxel,
vinblastine, vincristine, cyclophosphamide, apigenin, genistein,
cycloxazoline.
The compounds described herein may also be used in combination with
substances which inhibit signal transduction mediated by human VEGF
receptor Flt-1.
The compounds described herein may also be used for treating or
preventing a tumor necrosis factor-mediated disease comprising
co-administering a tumor necrosis factor alpha antagonist and an
effective amount of a compound of Formula (I) to a patient.
Contemplated tumor necrosis factor-mediated diseases include but
are not limited to autoimmune disease, acute or chronic immune
disease, inflammatory disease and neurodegenerative disease.
This invention further relates to a method for inhibiting abnormal
cell growth in a mammal which method comprises administering to the
mammal an amount of a compound of formula (I), or prodrugs thereof,
pharmaceutically active metabolites, pharmaceutically acceptable
salts, or pharmaceutically acceptable solvates of said compounds
and said prodrugs, in combination with radiation therapy, wherein
the amount of the compound, salt, solvate or prodrug is in
combination with the radiation therapy effective in inhibiting
abnormal cell growth in the mammal. Techniques for administering
radiation therapy are known in the art, and these techniques can be
used in the combination therapy described herein. The
administration of the compound of the invention in this combination
therapy can be determined as described herein.
It is believed that the compounds of formula (I) can render
abnormal cells more sensitive to treatment with radiation for
purposes of killing and/or inhibiting the growth of such cells.
Accordingly, this invention further relates to a method for
sensitizing abnormal cells in a mammal to treatment with radiation
which comprises administering to the mammal an amount of a compound
of formula (I), or prodrugs thereof, pharmaceutically active
metabolites, pharmaceutically acceptable salts, or pharmaceutically
acceptable solvates of said compounds and said prodrugs, which
amount is effective in sensitizing abnormal cells to or enhancing
the effects of treatment with radiation. The amount of the
compound, salt, solvate or prodrug of formula (I) in this method
can be determined according to the means for ascertaining effective
amounts of such compounds described herein.
This invention further relates to a method for treating a disease
related to vasculogenesis or angiogenesis in a mammal comprising
administering to said mammal a therapeutically effective amount of
a compound of formula (I), or prodrugs thereof, pharmaceutically
active metabolites, pharmaceutically acceptable salts, or
pharmaceutically acceptable solvates of said compounds and said
prodrugs, in conjunction with a therapeutically effective amount of
an anti-hypertensive agent.
Compounds of the present invention may be used in combination with
CHK-1 inhibitors. Certain CHK-1 inhibitors have been proposed for
cancer therapy (see Sanchez, Y. et.al. (1997) Science 277: 1497
1501 and Flaggs, G. et. al. (1997) Current Biology 7:977 986; U.S.
Pat. Nos. 6,413,755, 6,383,744, and 6,211,164; and International
Publication Nos. WO 01/16306, WO 01/21771, WO 00/16781, and WO
02/070494). In this embodiment, the CHK-1 inhibitor may be
administered as a single agent or as co-therapy with other
anti-neoplasm therapies including anti-neoplastic agents and
radiation therapy.
The wide variety of available anti-neoplastic agents are
contemplated for combination therapy with CHK-1 in accordance with
present invention. In a preferred embodiment, anti-neoplastic
agents that assert their cytotoxic effects by activating programmed
cell death or apoptosis may be used in combination with the CHK-1
inhibitor. The anti-neoplastic agents contemplated in accordance
with the present invention include, but are not limited to
alkylating agents, including busulfan, chlorambucil,
cyclophosphamide, iphosphamide, melphalan, nitrogen mustard,
streptozocin, thiotepa, uracil nitrogen mustard,
triethylenemelamine, temozolomide, and SARCnu; antibiotics and
plant alkaloids including actinomycin-D, bleomycin, cryptophycins,
daunorubicin, doxorubicin, idarubicin, irinotecan, L-asparaginase,
mitomycin-C, mitramycin, navelbine, paclitaxel, docetaxel,
topotecan, vinblastine, vincristine, VM-26, and VP-16-213; hormones
and steroids including 5.alpha.-reductase inhibitor,
aminoglutethimide, anastrozole, bicalutamide, chlorotrianisene,
DES, dromostanolone, estramustine, ethinyl estradiol, flutamide,
fluoxymesterone, goserelin, hydroxyprogesterone, letrozole,
leuprolide, medroxyprogesterone acetate, megestrol acetate, methyl
prednisolone, methyltestosterone, mitotane, nilutamide,
prednisolone, SERM3, tamoxifen, testolactone, testosterone,
triamicnolone, and zoladex; synthetics including all-trans retinoic
acid, BCNU (carmustine), CBDCA carboplatin (paraplatin), CCNU
(lomustine), cis-diaminedichloroplatinum (cisplatin), dacarbazine,
gliadel, hexamethylmelamine, hydroxyurea, levamisole, mitoxantrone,
o, p'-DDD (lysodren, mitotane), oxaliplatin, porfimer sodium,
procarbazine, GleeVec; antimetabolites including
chlorodeoxyadenosine, cytosine arabinoside, 2'-deoxycoformycin,
fludarabine phosphate, 5-fluorouracil, 5-FUDR, gemcitabine,
camptothecin, 6-mercaptopurine, methotrexate, MTA, and thioguanine;
and biologics including alpha interferon, BCG, G-CSF, GM-CSF,
interleukin-2, herceptin; and the like.
In a preferred embodiment of the invention, the anti-neoplastic
agent is selected from the group consisting of alkylating agents,
antibiotics and plant alkaloids, hormones and steroids, synthetic
agents having anti-neoplastic activity, antimetabolites and
biological molecules having anti-neoplastic activity.
In a preferred embodiment of the invention the antineoplastic agent
is selected from the group consisting of Ara-c, VP-16, cis-platin,
adriamycin, 2-chloro-2-deoxyadenosine,
9-.beta.-D-arabinosyl-2-fluoroadenine, carboplatin, gemcitabine,
camptothecin, paclitaxel, BCNU, 5-fluorouracil, irinotecan, and
doxorubicin; more preferably gemcitabine.
The CHK-1 inhibitor in combination with the VEGF inhibitor
identified in the present invention may also enhance the
antineoplasm effects of radiation therapy. Usually, radiation can
be used to treat the site of a solid tumor directly or administered
by brachytherapy implants. The various types of therapeutic
radiation which are contemplated for combination therapy in
accordance with the present invention may be those used in the
treatment of cancer which include, but are not limited to X-rays,
gamma radiation, high energy electrons and High LET (Linear Energy
Transfer) radiation such as protons, neutrons, and alpha particles.
The ionizing radiation may be employed by techniques well known to
those skilled in the art. For example, X-rays and gamma rays are
applied by external and/or interstitial means from linear
accelerators or radioactive sources. High-energy electrons may be
produced by linear accelerators. High LET radiation is also applied
from radioactive sources implanted interstitially.
The compounds of formula (I) or prodrugs thereof, pharmaceutically
active metabolites, pharmaceutically acceptable salts, or
pharmaceutically acceptable solvates of said compounds and said
prodrugs, can each independently also be used in a palliative
neo-adjuvant/adjuvant therapy in alleviating the symptoms
associated with the diseases recited herein as well as the symptoms
associated with abnormal cell growth. Such therapy can be a
monotherapy or can be in a combination with chemotherapy and/or
immunotherapy.
If the substituents themselves are not compatible with the
synthetic methods of this invention, the substituent may be
protected with a suitable protecting group that is stable to the
reaction conditions used in these methods. The protecting group may
be removed at a suitable point in the reaction sequence of the
method to provide a desired intermediate or target compound.
Suitable protecting groups and the methods for protecting and
de-protecting different substituents using such suitable protecting
groups are well known to those skilled in the art; examples of
which may be found in T. Greene and P. Wuts, Protecting Groups in
Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999),
which is incorporated herein by reference in its entirety. In some
instances, a substituent may be specifically selected to be
reactive under the reaction conditions used in the methods of this
invention. Under these circumstances, the reaction conditions
convert the selected substituent into another substituent that is
either useful in an intermediate compound in the methods of this
invention or is a desired substituent in a target compound.
The compounds of the present invention may have asymmetric carbon
atoms. Such diastereomeric mixtures can be separated into their
individual diastereomers on the basis of their physical chemical
differences by methods known to those skilled in the art, for
example, by chromatography or fractional crystallization.
Enantiomers can be separated by converting the enantiomeric
mixtures into a diastereomeric mixture by reaction with an
appropriate optically active compound (e.g., alcohol), separating
the diastereomers and converting (e.g., hydrolyzing) the individual
diastereomers to the corresponding pure enantiomers. All such
isomers, including diastereomeric mixtures and pure enantiomers are
considered as part of the invention.
The compounds of present invention may in certain instances exist
as tautomers. This invention relates to the use of all such
tautomers and mixtures thereof.
Preferably, the compounds of the present invention are used in a
form that is at least 90% optically pure, that is, a form that
contains at least 90% of a single isomer (80% enantiomeric excess
("e.e.") or diastereomeric excess ("d.e.")), more preferably at
least 95% (90% e.e. or d.e.), even more preferably at least 97.5%
(95% e.e. or d.e.), and most preferably at least 99% (98% e.e. or
d.e.).
Additionally, the formulae are intended to cover solvated as well
as unsolvated forms of the identified structures. For example,
Formula I includes compounds of the indicated structure in both
hydrated and non-hydrated forms. Additional examples of solvates
include the structures in combination with isopropanol, ethanol,
methanol, DMSO, ethyl acetate, acetic acid, or ethanolamine.
In the case of agents that are solids, it is understood by those
skilled in the art that the inventive compounds and salts may exist
in different crystal or polymorphic forms, all of which are
intended to be within the scope of the present invention and
specified formulas.
This invention also encompasses pharmaceutical compositions
containing and methods of treating bacterial infections through
administering prodrugs of compounds of the formula 1. Compounds of
formula 1 having free amino, amido, hydroxy or carboxylic groups
can be converted into prodrugs. Prodrugs include compounds wherein
an amino acid residue, or a polypeptide chain of two or more (e.g.,
two, three or four) amino acid residues is covalently joined
through an amide or ester bond to a free amino, hydroxy or
carboxylic acid group of compounds of formula 1. The amino acid
residues include but are not limited to the 20 naturally occurring
amino acids commonly designated by three letter symbols and also
includes 4-hydroxyproline, hydroxylysine, demosine, isodemosine,
3-methylhistidine, norvalin, beta-alanine, gamma-aminobutyric acid,
citrulline homocysteine, homoserine, ornithine and methionine
sulfone. Additional types of prodrugs are also encompassed. For
instance, free carboxyl groups can be derivatized as amides or
alkyl esters. Free hydroxy groups may be derivatized using groups
including but not limited to hemisuccinates, phosphate esters,
dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as
outlined in Advanced Drug Delivery Reviews, 1996, 19, 115.
Carbamate prodrugs of hydroxy and amino groups are also included,
as are carbonate prodrugs, sulfonate esters and sulfate esters of
hydroxy groups. Derivatization of hydroxy groups as (acyloxy)methyl
and (acyloxy)ethyl ethers wherein the acyl group may be an alkyl
ester, optionally substituted with groups including but not limited
to ether, amine and carboxylic acid functionalities, or where the
acyl group is an amino acid ester as described above, are also
encompassed. Prodrugs of this type are described in J. Med. Chem.
1996, 39, 10. Free amines can also be derivatized as amides,
sulfonamides or phosphonamides. All of these prodrug moieties may
incorporate groups including but not limited to ether, amine and
carboxylic acid functionalities.
Definitions
As used herein, the following terms have the following meanings,
unless expressly indicated otherwise.
The term "comprising" and "including" are used in their open,
non-limiting sense.
The terms "abnormal cell growth" and "hyperproliferative disorder"
are used interchangeably in this application.
"Abnormal cell growth" refers to cell growth that is independent of
normal regulatory mechanisms (e.g., loss of contact inhibition),
including the abnormal growth of normal cells and the growth of
abnormal cells. This includes, but is not limited to, the abnormal
growth of: (1) tumor cells (tumors), both benign and malignant,
expressing an activated Ras oncogene; (2) tumor cells, both benign
and malignant, in which the Ras protein is activated as a result of
oncogenic mutation in another gene; (3) benign and malignant cells
of other proliferative diseases in which aberrant Ras activation
occurs. Examples of such benign proliferative diseases are
psoriasis, benign prostatic hypertrophy, human papilloma virus
(HPV), and restinosis. "Abnormal cell growth" also refers to and
includes the abnormal growth of cells, both benign and malignant,
resulting from activity of the enzyme farnesyl protein
transferase.
The term "acyl" includes alkyl, aryl, or heteroaryl substituents
attached to a compound via a carbonyl functionality (e.g.,
--C(O)-alkyl, --C(O)-aryl, etc.).
The term "acylamino" refers to an acyl radical appended to an amino
or alkylamino group, and includes --C(O)--NH.sub.2 and
--C(O)--NRR'' groups where R and R' are as defined in conjunction
with alkylamino.
The term "acyloxy" refers to the ester group --OC(O)--R, where R is
H, alkyl, alkenyl, alkynyl, or aryl.
##STR00013## where each of R and R' are independently selected from
the group consisting of H, alkyl, and aryl.
The term "alkenyl" includes alkyl moieties having at least one
carbon-carbon double bond, including E and Z isomers of said
alkenyl moiety. The term also includes cycloalkyl moieties having
at least one carbon-carbon double bond, i.e., cycloalkenyl.
Examples of alkenyl radicals include ethenyl, propenyl, butenyl,
1,4-butadienyl, cyclopentenyl, cyclohexenyl, prop-2-enyl,
but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl, and the
like. An alkenyl group may be optionally substituted.
The term "alkenylene" refers to a divalent straight chain, branched
chain or cyclic saturated aliphatic group containing at least one
carbon-carbon double bond, and including E and Z isomers of said
alkenylene moiety. An alkyenylene group may be optionally
substituted.
The term "alkoxy" means an O-alkyl group. Examples of alkoxy
radicals include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy,
iso-butoxy, sec-butoxy, tert-butoxy and the like.
The term "alkyl" means saturated monovalent hydrocarbon radicals
having straight, cyclic or branched moieties. An "alkyl" group may
include an optional carbon-carbon double or triple bond where the
alkyl group comprises at least two carbon atoms. Cycloalkyl
moieties require at least three carbon atoms. Examples of straight
or branched alkyl radicals include methyl (Me), ethyl (Et),
n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl,
tert-amyl, pentyl, isopentyl, hexyl, heptyl, octyl and the like. An
alkyl group may be optionally substituted.
The term "alkylamino" refers to the --NRR' group, where R and R'
are independently selected from hydrogen (however, R and R' cannot
both be hydrogen), alkyl, and aryl groups; or R and R', taken
together, can form a cyclic ring system.
The term "alkylene" refers to a divalent straight chain, branched
chain or cyclic saturated aliphatic group. The latter group may
also be referred to more specifically as a cycloalkylene group. An
alkylene group may be optionally substituted.
The term "alkylthio" alone or in combination, refers to an
optionally substituted alkyl thio radical, alkyl-S--.
The term "alkynyl" refers to straight- and branched-chain alkynyl
groups having from two to twelve carbon atoms, preferably from 2 to
6 carbons, and more preferably from 2 to 4 carbons. Illustrative
alkynyl groups include prop-2-ynyl, but-2-ynyl, but-3-ynyl,
2-methylbut-2-ynyl, hex-2-ynyl, and the like. An alkynyl group may
be optionally substituted.
The term "amide" refers to the radical --C(O)N(R')(R'') where R'
and R'' are each independently selected from hydrogen, alkyl,
alkenyl, alkynyl, --OH, alkoxy, cycloalkyl, heterocycloalkyl,
heteroaryl, aryl as defined above; or R' and R'' cyclize together
with the nitrogen to form a heterocycloalkyl or heteroaryl.
The term "amino" refers to the --NH.sub.2 group.
The term "anti-neoplastic agent" refers to agents capable of
inhibiting or preventing the growth of neoplasms, or checking the
maturation and proliferation of malignant (cancer) cells.
The term "aromatic" refers to compounds or moieties comprising
multiple conjugated double bonds. Examples of aromatic moieties
include, without limitation, aryl or heteroaryl ring systems.
The term "aryl" (Ar) means an organic radical derived from a
monocyclic or polycyclic aromatic hydrocarbon by removal of one
hydrogen, such as phenyl or naphthyl. Preferred aryl groups have
from 4 to 20 ring atoms, and more preferably from 6 to 14 ring
atoms. An aryl group may be optionally substituted. Illustrative
examples of aryl groups include the following moieties:
##STR00014## and the like.
The term "aryloxy" means aryl-O--.
The term "arylthio" means an aryl thio radical, aryl-S--.
The term "carbamoyl" or "carbamate" refers to the group
--O--C(O)--NRR'' where R and R'' are independently selected from
hydrogen, alkyl, and aryl groups; and R and R'' taken together can
form a cyclic ring system.
The term "carbocycle" includes optionally substituted cycloalkyl
and aryl moieties. The term "carbocycle" also includes cycloalkenyl
moieties having at least one carbon-carbon double bond.
The term "carboxy esters" refers to --C(O)OR where R is alkyl or
aryl.
The term "cycloalkyl" refers to a monocyclic or polycyclic radical
which contains only carbon and hydrogen, and may be saturated,
partially unsaturated, or fully unsaturated. A cycloalkyl group may
be optionally substituted. Preferred cycloalkyl groups include
groups having from three to twelve ring atoms, more preferably from
5 to 10 ring atoms. Illustrative examples of cycloalkyl groups
include the following moieties:
##STR00015## and compounds of the like.
The term "halo" or "halogen" means fluoro, chloro, bromo or iodo.
Preferred halo groups are fluoro, chloro and bromo.
The terms haloalkyl, haloalkenyl, haloalkynyl and haloalkoxy
include alkyl, alkenyl, alkynyl and alkoxy structures, that are
substituted with one or more halo groups or with combinations
thereof.
The terms "heteroalkyl" "heteroalkenyl" and "heteroalkynyl" include
optionally substituted alkyl, alkenyl and alkynyl radicals and
which have one or more skeletal chain atoms selected from an atom
other that carbon, e.g., oxygen, nitrogen, sulfur, phosphorus or
combinations thereof.
The term "heteroaryl" (heteroAr) refers to an aryl group that
includes one or more ring heteroatoms selected from nitrogen,
oxygen and sulfur. A heteroaryl group may be optionally
substituted. The polycyclic heteroaryl group may be fused or
non-fused. Illustrative examples of aryl groups include the
following moieties:
##STR00016## and the like.
The term "heterocycle" refers to aromatic and non-aromatic
heterocyclic groups containing one to four heteroatoms each
selected from O, S and N, wherein each heterocyclic group has from
4 to 10 atoms in its ring system, and with the proviso that the
ring of said group does not contain two adjacent O or S atoms.
Non-aromatic heterocyclic groups include groups having only 4 atoms
in their ring system, but aromatic heterocyclic groups must have at
least 5 atoms in their ring system. The heterocyclic groups include
benzo-fused ring systems. An example of a 4 membered heterocyclic
group is azetidinyl (derived from azetidine). An example of a 5
membered heterocyclic group is thiazolyl. An example of a 6
membered heterocyclic group is pyridyl, and an example of a 10
membered heterocyclic group is quinolinyl. Examples of non-aromatic
heterocyclic groups are pyrrolidinyl, tetrahydrofuranyl,
dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl,
dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino,
thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl,
thietanyl, homopiperidinyl, oxepanyl, thiepanyl, oxazepinyl,
diazepinyl, thiazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl,
3-pyrrolinyl, indolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl,
1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl,
dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl,
imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl,
3-azabicyclo[4.1.0]heptanyl, 3H-indolyl and quinolizinyl. Examples
of aromatic heterocyclic groups are pyridinyl, imidazolyl,
pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl,
thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl,
quinolinyl, isoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,
cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,
triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl,
thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,
benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,
naphthyridinyl, and furopyridinyl. The foregoing groups, as derived
from the groups listed above, may be C-attached or N-attached where
such is possible. For instance, a group derived from pyrrole may be
pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a
group derived from imidazole may be imidazol-1-yl or imidazol-3-yl
(both N-attached) or imidazol-2-yl, imidazol-4-yl or imidazol-5-yl
(all C-attached). The heterocyclic groups include benzo-fused ring
systems and ring systems substituted with one or two oxo (.dbd.O)
moieties such as pyrrolidin-2-one. A heterocycle group may be
optionally substituted.
The term "heterocyclic" comprises both heterocycloalkyl and
heteroaryl groups.
A "heterocycloalkyl" group refers to a cycloalkyl group that
includes at least one heteroatom selected from nitrogen, oxygen and
sulfur. The radicals may be fused with an aryl or heteroaryl.
Illustrative examples of heterocycloalkyl groups include
##STR00017## and the like.
The terms "5 membered heterocyclic", "5 or 6 membered
heterocyclic", "5 to 8 membered heterocyclic", "5 to 10 membered
heterocyclic" or "5 to 13 membered heterocyclic" includes aromatic
and non-aromatic heterocyclic groups containing one to four
heteroatoms each selected from O, S and N, wherein each
heterocyclic group has from 5, 6, 5 to 8, 5 to 10 or 5 to 13 atoms
in its ring system, respectively.
The term "membered ring" can embrace any cyclic structure. The term
"membered" is meant to denote the number of skeletal atoms that
constitute the ring. Thus, for example, cyclohexyl, pyridine, pyran
and thiopyran are 6-membered rings and cyclopentyl, pyrrole, furan,
and thiophene are 5-membered rings.
The term "neoplasm" is defined as in Stedman's Medical Dictionary
25.sup.th Edition (1990)and refers to an abnormal tissue that grows
by cellular proliferation more rapidly than normal and continues to
grow after the stimuli that initiated the new growth ceases.
Neoplasms show partial or complete lack of structural organization
and functional coordination compared with normal tissue, and
usually form a distinct mass of tissue that may be either benign
(benign tumor) or malignant (cancer).
"Optionally substituted" groups may be substituted or
unsubstituted. When substituted, the substituents of an "optionally
substituted" group may include, without limitation, one or more
substituents independently selected from the following groups or
designated subsets thereof: (C.sub.1 C.sub.6)alkyl, (C.sub.2
C.sub.6)alkenyl, (C.sub.2 C.sub.6)alkynyl, (C.sub.1
C.sub.6)heteroalkyl, (C.sub.1 C.sub.6)haloalkyl, (C.sub.2
C.sub.6)haloalkenyl, (C.sub.2 C.sub.6)haloalkynyl, (C.sub.3
C.sub.6)cycloalkyl, phenyl, (C.sub.1 C.sub.6)alkoxy, phenoxy,
(C.sub.1 C.sub.6)haloalkoxy, amino, (C.sub.1 C.sub.6)alkylamino,
(C.sub.1 C.sub.6)alkylthio, phenyl-S--, oxo, (C.sub.1
C.sub.6)carboxyester, (C.sub.1 C.sub.6)carboxamido, (C.sub.1
C.sub.6)acyloxy, H, halogen, CN, NO.sub.2, NH.sub.2, N.sub.3,
NHCH.sub.3, N(CH.sub.3).sub.2, SH, SCH.sub.3, OH, OCH.sub.3,
OCF.sub.3, CH.sub.3, CF.sub.3, C(O)CH.sub.3, CO.sub.2CH.sub.3,
CO.sub.2H, C(O)NH.sub.2, pyridinyl, thiophene, furanyl, (C.sub.1
C.sub.6)carbamate, and (C.sub.1 C.sub.6)urea. An optionally
substituted group may be unsubstituted (e.g., --CH.sub.2CH.sub.3),
fully substituted (e.g., --CF.sub.2CF.sub.3), monosubstituted
(e.g., --CH.sub.2CH.sub.2F) or substituted at a level anywhere
in-between fully substituted and monosubstituted (e.g.,
--CH.sub.2CF.sub.3).
The term "oxo" means an "O" group.
The term "perhalo" refers to groups wherein every C--H bond has
been replaced with a C-halo bond on an aliphatic or aryl group.
Examples of perhaloalkyl groups include --CF.sub.3 and
--CFCl.sub.2.
The term "substituted" means that the group in question, e.g.,
alkyl group, etc., may bear one or more substituents.
The term "ureyl" or "urea" refers to the group --N(R)--C(O)--NR'R''
where R, R', and R'' are independently selected from hydrogen,
alkyl, aryl; and where each of R--R', R'R'', or R--R'' taken
together can form a cyclic ring system.
Pharmaceutical Formulations and Compositions
In addition to compounds of Formula I, the invention includes
N-oxides, pharmaceutically acceptable prodrugs, pharmaceutically
acceptable solvates, pharmaceutically active metabolites, and
pharmaceutically acceptable salts of such compounds, prodrugs,
solvates and metabolites.
The term "pharmaceutically acceptable" means pharmacologically
acceptable and substantially non-toxic to the subject being
administered the agent.
A "pharmacological composition" refers to a mixture of one or more
of the compounds described herein, or physiologically acceptable
salts thereof, with other chemical components, such as
physiologically acceptable carriers and/or excipients. The purpose
of a pharmacological composition is to facilitate administration of
a compound to an organism.
A "physiologically acceptable carrier" refers to a carrier or
diluent that does not cause significant or otherwise unacceptable
irritation to an organism and does not unacceptably abrogate the
biological activity and properties of the administered
compound.
An "excipient" generally refers to substance, often an inert
substance, added to a pharmacological composition or otherwise used
as a vehicle to further facilitate administration of a compound.
Examples of excipients include but are not limited to calcium
carbonate, calcium phosphate, various sugars and types of starch,
cellulose derivatives, gelatin, vegetable oils and polyethylene
glycols.
The term "prodrug" means compounds that are drug precursors, which
following administration, release the drug in vivo via some
chemical or physiological process (e.g., a prodrug on being brought
to the physiological pH is converted to the desired drug form).
Prodrugs include compounds wherein an amino acid residue, or a
polypeptide chain of two or more (e.g., two, three or four) amino
acid residues is covalently joined through an amide or ester bond
to a free amino, hydroxy or carboxylic acid group of compounds of
formula (I). The amino acid residues include but are not limited to
the 20 naturally occurring amino acids commonly designated by three
letter symbols and also includes 4-hydroxyproline, hydroxylysine,
demosine, isodemosine, 3-methylhistidine, norvalin, beta-alanine,
gamma-aminobutyric acid, citrulline homocysteine, homoserine,
ornithine and methionine sulfone. Additional types of prodrugs are
also encompassed. For instance, free carboxyl groups can be
derivatized as amides or alkyl esters. Free hydroxy groups may be
derivatized using groups including but not limited to
hemisuccinates, phosphate esters, dimethylaminoacetates, and
phosphoryloxymethyloxycarbonyls, as outlined in Advanced Drug
Delivery Reviews, 1996, 19, 115. Carbamate prodrugs of hydroxy and
amino groups are also included, as are carbonate prodrugs,
sulfonate esters and sulfate esters of hydroxy groups.
Derivatization of hydroxy groups as (acyloxy)methyl and
(acyloxy)ethyl ethers wherein the acyl group may be an alkyl ester,
optionally substituted with groups including but not limited to
ether, amine and carboxylic acid functionalities, or where the acyl
group is an amino acid ester as described above, are also
encompassed. Prodrugs of this type are described in J. Med. Chem.
1996, 39, 10. Free amines can also be derivatized as amides,
sulfonamides or phosphonamides. All of these prodrug moieties may
incorporate groups including but not limited to ether, amine and
carboxylic acid functionalities.
"A pharmaceutically acceptable prodrug" is a compound that may be
converted under physiological conditions or by solvolysis to the
specified compound or to a pharmaceutically acceptable salt of such
compound. "A pharmaceutically active metabolite" is intended to
mean a pharmacologically active product produced through metabolism
in the body of a specified compound or salt thereof. Prodrugs and
active metabolites of a compound may be identified using routine
techniques known in the art. See, e.g., Bertolini et al., J. Med.
Chem., 40, 2011 2016 (1997); Shan et al., J. Pharm. Sci., 86 (7),
765 767; Bagshawe, Drug Dev. Res., 34, 220 230 (1995); Bodor,
Advances in Drug Res., 13, 224 331 (1984); Bundgaard, Design of
Prodrugs (Elsevier Press 1985); and Larsen, Design and Application
of Prodrugs, Drug Design and Development (Krogsgaard-Larsen et al.,
eds., Harwood Academic Publishers, 1991).
"A pharmaceutically acceptable salt" is intended to mean a salt
that retains the biological effectiveness of the free acids and
bases of the specified compound and that is not biologically or
otherwise undesirable. A compound of the invention may possess a
sufficiently acidic, a sufficiently basic, or both functional
groups, and accordingly react with any of a number of inorganic or
organic bases, and inorganic and organic acids, to form a
pharmaceutically acceptable salt. Exemplary pharmaceutically
acceptable salts include those salts prepared by reaction of the
compounds of the present invention with a mineral or organic acid
or an inorganic base, such as salts including sulfates,
pyrosulfates, bisulfates, sulfites, bisulfites, phosphates,
monohydrogenphosphates, dihydrogenphosphates, metaphosphates,
pyrophosphates, chlorides, bromides, iodides, acetates,
propionates, decanoates, caprylates, acrylates, formates,
isobutyrates, caproates, heptanoates, propiolates, oxalates,
malonates, succinates, suberates, sebacates, fumarates, maleates,
butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates,
methylbenzoates, dinitrobenzoates, hydroxybenzoates,
methoxybenzoates, phthalates, sulfonates, xylenesulfonates,
phenylacetates, phenylpropionates, phenylbutyrates, citrates,
lactates, .gamma.-hydroxybutyrates, glycolates, tartrates,
methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates,
naphthalene-2-sulfonates, and mandelates.
If the compound of the invention is a base, the desired
pharmaceutically acceptable salt may be prepared by any suitable
method available in the art, for example, treatment of the free
base with an inorganic acid, such as hydrochloric acid, hydrobromic
acid, sulfuric acid, sulfamic acid, nitric acid, phosphoric acid
and the like, or with an organic acid, such as acetic acid,
phenylacetic acid, propionic acid, stearic acid, lactic acid,
ascorbic acid, maleic acid, hydroxymaleic acid, isethionic acid,
succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic
acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl
acid, such as glucuronic acid or galacturonic acid, an
alpha-hydroxy acid, such as citric acid or tartaric acid, an amino
acid, such as aspartic acid or glutamic acid, an aromatic acid,
such as benzoic acid, 2-acetoxybenzoic acid or cinnamic acid, a
sulfonic acid, such as p-toluenesulfonic acid, methanesulfonic acid
or ethanesulfonic acid, or the like.
If the compound of the invention is an acid, the desired
pharmaceutically acceptable salt may be prepared by any suitable
method, for example, treatment of the free acid with an inorganic
or organic base, such as an amine (primary, secondary or tertiary),
an alkali metal hydroxide or alkaline earth metal hydroxide, or the
like. Illustrative examples of suitable salts include organic salts
derived from amino acids, such as glycine and arginine, ammonia,
carbonates, bicarbonates, primary, secondary, and tertiary amines,
and cyclic amines, such as benzylamines, pyrrolidines, piperidine,
morpholine and piperazine, and inorganic salts derived from sodium,
calcium, potassium, magnesium, manganese, iron, copper, zinc,
aluminum and lithium.
Pharmaceutical compositions according to the invention may,
alternatively or in addition to a compound of Formula (I), comprise
as an active ingredient pharmaceutically acceptable prodrugs,
pharmaceutically active metabolites, and pharmaceutically
acceptable salts of such compounds and metabolites. Such compounds,
prodrugs, multimers, salts, and metabolites are sometimes referred
to herein collectively as "active agents" or "agents."
It will be appreciated that any solvate (e.g. hydrate) form of
compounds of formula (I) and prodrugs thereof can be used for the
purpose of the present invention.
Therapeutically effective amounts of the active agents of the
invention may be used to treat diseases mediated by modulation or
regulation of protein kinases. An "effective amount" is intended to
mean that amount of an agent that significantly inhibits
proliferation and/or prevents de-differentiation of a eukaryotic
cell, e.g., a mammalian, insect, plant or fungal cell, and is
effective for the indicated utility, e.g., specific therapeutic
treatment.
The compositions containing the compound(s) of the described herein
can be administered for prophylactic and/or therapeutic treatments.
In therapeutic applications, the compositions are administered to a
patient already suffering from a proliferative disorder or
condition (including, but not limited to, cancer), as described
above, in an amount sufficient to cure or at least partially arrest
the symptoms of the proliferative disorder or condition. An amount
adequate to accomplish this is defined as "therapeutically
effective amount or dose."Amounts effective for this use will
depend on the severity and course of the proliferative disorder or
condition, previous therapy, the patient's health status and
response to the drugs, and the judgment of the treating physician.
In prophylactic applications, compositions containing the compounds
described herein are administered to a patient susceptible to or
otherwise at risk of a particular proliferative disorder or
condition. Such an amount is defined to be a "prophylactically
effective amount or dose." In this use, the precise amounts also
depend on the patient's state of health, weight, and the like. It
is considered well within the skill of the art for one to determine
such therapeutically effective or prophylactically effective
amounts by routine experimentation (e.g., a dose escalation
clinical trial).
The terms "enhance" or "enhancing" means to increase or prolong
either in potency or duration a desired effect. Thus, in regard to
enhancing the effect of therapeutic agents, the term "enhancing"
refers to the ability to increase or prolong, either in potency or
duration, the effect of other therapeutic agents on a system (e.g.,
a tumor cell). An "enhancing-effective amount," as used herein,
refers to an amount adequate to enhance the effect of another
therapeutic agent in a desired system (including, by way of example
only, a tumor cell in a patient). When used in a patient, amounts
effective for this use will depend on the severity and course of
the proliferative disorder (including, but not limited to, cancer),
previous therapy, the patient's health status and response to the
drugs, and the judgment of the treating physician. It is considered
well within the skill of the art for one to determine such
enhancing-effective amounts by routine experimentation.
Once improvement of the patient's conditions has occurred, a
maintenance dose is administered if necessary. Subsequently, the
dosage or the frequency of administration, or both, can be reduced,
as a function of the symptoms, to a level at which the improved
proliferative disorder or condition is retained. When the symptoms
have been alleviated to the desired level, treatment can cease.
Patients can, however, require intermittent treatment on a
long-term basis upon any recurrence of the disease symptoms.
The amount of a given agent that will correspond to such an amount
will vary depending upon factors such as the particular compound,
disease condition and its severity, the identity (e.g., weight) of
the subject or host in need of treatment, but can nevertheless be
routinely determined in a manner known in the art according to the
particular circumstances surrounding the case, including, e.g., the
specific agent being administered, the route of administration, the
condition being treated, and the subject or host being treated.
"Treating" is intended to mean at least the mitigation of a disease
condition in a subject such as mammal (e.g., human), that is
affected, at least in part, by the activity of one or more kinases,
for example protein kinases such as tyrosine kinases, and includes:
preventing the disease condition from occurring in a mammal,
particularly when the mammal is found to be predisposed to having
the disease condition but has not yet been diagnosed as having it;
modulating and/or inhibiting the disease condition; and/or
alleviating the disease condition.
Agents that potently regulate, modulate, or inhibit cell
proliferation are preferred. For certain mechanisms, inhibition of
the protein kinase activity associated with CDK complexes, among
others, and those which inhibit angiogenesis and/or inflammation
are preferred. The present invention is further directed to methods
of modulating or inhibiting protein kinase activity, for example in
mammalian tissue, by administering a compound of Formula (I). The
activity of agents as anti-proliferatives is easily measured by
known methods, for example by using whole cell cultures in an MTT
assay. The activity of the compounds of Formula (I) as modulators
of protein kinase activity, such as the activity of kinases, may be
measured by any of the methods available to those skilled in the
art, including in vivo and/or in vitro assays. Examples of suitable
assays for activity measurements include those described in
International Publication No. WO 99/21845; Parast et al.,
Biochemistry, 37, 16788 16801 (1998); Connell-Crowley and Harpes,
Cell Cycle: Materials and Methods, (Michele Pagano, ed. Springer,
Berlin, Germany)(1995); International Publication No. WO 97/34876;
and International Publication No. WO 96/14843. These properties may
be assessed, for example, by using one or more of the biological
testing procedures set out in the examples below.
The active agents of the invention may be formulated into
pharmaceutical compositions as described below. Pharmaceutical
compositions of this invention comprise an effective modulating,
regulating, or inhibiting amount of a compound of Formula I and an
inert, pharmaceutically acceptable carrier or diluent. In one
embodiment of the pharmaceutical compositions, efficacious levels
of the compounds of Formula (I) are provided so as to provide
therapeutic benefits involving anti-proliferative ability. By
"efficacious levels" is meant levels in which proliferation is
inhibited, or controlled. These compositions are prepared in
unit-dosage form appropriate for the mode of administration, e.g.,
parenteral or oral administration.
A compound of Formula (I) can be administered in conventional
dosage form prepared by combining a therapeutically effective
amount of an agent (e.g., a compound of Formula I) as an active
ingredient with appropriate pharmaceutical carriers or diluents
according to conventional procedures. These procedures may involve
mixing, granulating and compressing or dissolving the ingredients
as appropriate to the desired preparation.
The pharmaceutical carrier employed may be either a solid or
liquid. Exemplary of solid carriers are lactose, sucrose, talc,
gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and
the like. Exemplary of liquid carriers are syrup, peanut oil, olive
oil, water and the like. Similarly, the carrier or diluent may
include time-delay or time-release material known in the art, such
as glyceryl monostearate or glyceryl distearate alone or with a
wax, ethylcellulose, hydroxypropylmethylcellulose,
methylmethacrylate and the like.
A variety of pharmaceutical forms can be employed. Thus, if a solid
carrier is used, the preparation can be tableted, placed in a hard
gelatin capsule in powder or pellet form or in the form of a troche
or lozenge. The amount of solid carrier may vary, but generally
will be from about 25 mg to about 1 g. If a liquid carrier is used,
the preparation will be in the form of syrup, emulsion, soft
gelatin capsule, sterile injectable solution or suspension in an
ampoule or vial or non-aqueous liquid suspension.
To obtain a stable water-soluble dose form, a pharmaceutically
acceptable salt of a compound of Formula (I) can be dissolved in an
aqueous solution of an organic or inorganic acid, such as 0.3M
solution of succinic acid or citric acid. If a soluble salt form is
not available, the agent may be dissolved in a suitable cosolvent
or combinations of cosolvents. Examples of suitable cosolvents
include, but are not limited to, alcohol, propylene glycol,
polyethylene glycol 300, polysorbate 80, glycerin and the like in
concentrations ranging from 0 60% of the total volume. In an
exemplary embodiment, a compound of Formula I is dissolved in DMSO
and diluted with water. The composition may also be in the form of
a solution of a salt form of the active ingredient in an
appropriate aqueous vehicle such as water or isotonic saline or
dextrose solution.
It will be appreciated that the actual dosages of the agents used
in the compositions of this invention will vary according to the
particular complex being used, the particular composition
formulated, the mode of administration and the particular site,
host and disease being treated. Optimal dosages for a given set of
conditions can be ascertained by those skilled in the art using
conventional dosage-determination tests in view of the experimental
data for an agent. For oral administration, an exemplary daily dose
generally employed is from about 0.001 to about 1000 mg/kg of body
weight, with courses of treatment repeated at appropriate
intervals. Administration of prodrugs is typically dosed at weight
levels that are chemically equivalent to the weight levels of the
fully active form.
The compositions of the invention may be manufactured in manners
generally known for preparing pharmaceutical compositions, e.g.,
using conventional techniques such as mixing, dissolving,
granulating, dragee-making, levigating, emulsifying, encapsulating,
entrapping or lyophilizing. Pharmaceutical compositions may be
formulated in a conventional manner using one or more
physiologically acceptable carriers, which may be selected from
excipients and auxiliaries that facilitate processing of the active
compounds into preparations that can be used pharmaceutically.
Proper formulation is dependent upon the route of administration
chosen. For injection, the agents of the invention may be
formulated into aqueous solutions, preferably in physiologically
compatible buffers such as Hanks's solution, Ringers solution, or
physiological saline buffer. For transmucosal administration,
penetrants appropriate to the barrier to be permeated are used in
the formulation. Such penetrants are generally known in the
art.
For oral administration, the compounds can be formulated readily by
combining the compounds with pharmaceutically acceptable carriers
known in the art. Such carriers enable the compounds of the
invention to be formulated as tablets, pills, dragees, capsules,
liquids, gels, syrups, slurries, suspensions and the like, for oral
ingestion by a patient to be treated. Pharmaceutical preparations
for oral use can be obtained using a solid excipient in admixture
with the active ingredient (agent), optionally grinding the
resulting mixture, and processing the mixture of granules after
adding suitable auxiliaries, if desired, to obtain tablets or
dragee cores. Suitable excipients include: fillers such as sugars,
including lactose, sucrose, mannitol, or sorbitol; and cellulose
preparations, for example, maize starch, wheat starch, rice starch,
potato starch, gelatin, gum, methyl cellulose,
hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or
polyvinylpyrrolidone (PVP). If desired, disintegrating agents may
be added, such as crosslinked polyvinylpyrrolidone, agar, or
alginic acid or a salt thereof such as sodium alginate.
Dragee cores are provided with suitable coatings. For this purpose,
concentrated sugar solutions may be used, which may optionally
contain gum arabic, polyvinyl pyrrolidone, Carbopol gel,
polyethylene glycol, and/or titanium dioxide, lacquer solutions,
and suitable organic solvents or solvent mixtures. Dyestuffs or
pigments may be added to the tablets or dragee coatings for
identification or to characterize different combinations of
agents.
Pharmaceutical preparations that can be used orally include
push-fit capsules made of gelatin, as well as soft, sealed capsules
made of gelatin and a plasticizer, such as glycerol or sorbitol.
The push-fit capsules can contain the agents in admixture with
fillers such as lactose, binders such as starches, and/or
lubricants such as talc or magnesium stearate, and, optionally,
stabilizers. In soft capsules, the agents may be dissolved or
suspended in suitable liquids, such as fatty oils, liquid paraffin,
or liquid polyethylene glycols. In addition, stabilizers may be
added. All formulations for oral administration should be in
dosages suitable for such administration. For buccal
administration, the compositions take the form of tablets or
lozenges formulated in conventional manners.
For administration intranasally or by inhalation, the compounds for
use according to the present invention are conveniently delivered
in the form of an aerosol spray presentation from pressurized packs
or a nebuliser, with the use of a suitable propellant, e.g.,
dichlorodifluoromethane, trichlorofluoromethane,
dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In
the case of a pressurized aerosol the dosage unit may be determined
by providing a valve to deliver a metered amount. Capsules and
cartridges of gelatin for use in an inhaler or insufflator and the
like may be formulated containing a powder mix of the compound and
a suitable powder base such as lactose or starch.
The compounds may be formulated for parenteral administration by
injection, e.g., by bolus injection or continuous infusion.
Formulations for injection may be presented in unit-dosage form,
e.g., in ampoules or in multi-dose containers, with an added
preservative. The compositions may take such forms as suspensions,
solutions or emulsions in oily or aqueous vehicles, and may contain
formulatory agents such as suspending, stabilizing and/or
dispersing agents.
Pharmaceutical formulations for parenteral administration include
aqueous solutions of the agents in water-soluble form.
Additionally, suspensions of the agents may be prepared as
appropriate oily injection suspensions. Suitable lipophilic
solvents or vehicles include fatty oils such as sesame oil, or
synthetic fatty acid esters, such as ethyl oleate or triglycerides,
or liposomes. Aqueous injection suspensions may contain substances
that increase the viscosity of the suspension, such as sodium
carboxymethyl cellulose, sorbitol, or dextran. Optionally, the
suspension may also contain suitable stabilizers or agents that
increase the solubility of the compounds to allow for the
preparation of highly concentrated solutions.
For administration to the eye, the agent is delivered in a
pharmaceutically acceptable ophthalmic vehicle such that the
compound is maintained in contact with the ocular surface for a
sufficient time period to allow the compound to penetrate the
corneal and internal regions of the eye, for example, the anterior
chamber, posterior chamber, vitreous body, aqueous humor, vitreous
humor, cornea, iris/ciliary, lens, choroid/retina and sclera. The
pharmaceutically acceptable ophthalmic vehicle may be an ointment,
vegetable oil, or an encapsulating material. A compound of the
invention may also be injected directly into the vitreous and
aqueous humor.
Alternatively, the agents may be in powder form for constitution
with a suitable vehicle, e.g., sterile pyrogen-free water, before
use. The compounds may also be formulated in rectal compositions
such as suppositories or retention enemas, e.g, containing
conventional suppository bases such as cocoa butter or other
glycerides.
In addition to the formulations described above, the agents may
also be formulated as a depot preparation. Such long-acting
formulations may be administered by implantation (for example,
subcutaneously or intramuscularly) or by intramuscular injection.
Thus, for example, the compounds may be formulated with suitable
polymeric or hydrophobic materials (for example, as an emulsion in
an acceptable oil) or ion-exchange resins, or as sparingly soluble
derivatives, for example, as a sparingly soluble salt.
An exemplary pharmaceutical carrier for hydrophobic compounds is a
cosolvent system comprising benzyl alcohol, a nonpolar surfactant,
a water-miscible organic polymer, and an aqueous phase. The
cosolvent system may be a VPD co-solvent system. VPD is a solution
of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant
polysorbate 80, and 65% w/v polyethylene glycol 300, made up to
volume in absolute ethanol. The VPD co-solvent system (VPD:5W)
contains VPD diluted 1:1 with a 5% dextrose in water solution. This
co-solvent system dissolves hydrophobic compounds well, and itself
produces low toxicity upon systemic administration. Naturally, the
proportions of a co-solvent system may be varied considerably
without destroying its solubility and toxicity characteristics.
Furthermore, the identity of the co-solvent components may be
varied: for example, other low-toxicity nonpolar surfactants may be
used instead of polysorbate 80; the fraction size of polyethylene
glycol may be varied; other biocompatible polymers may replace
polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars
or polysaccharides may be substituted for dextrose.
Alternatively, other delivery systems for hydrophobic
pharmaceutical compounds may be employed. Liposomes and emulsions
are known examples of delivery vehicles or carriers for hydrophobic
drugs. Certain organic solvents such as dimethylsulfoxide also may
be employed, although usually at the cost of greater toxicity.
Additionally, the compounds may be delivered using a
sustained-release system, such as semi-permeable matrices of solid
hydrophobic polymers containing the therapeutic agent. Various
sustained-release materials have been established and are known by
those skilled in the art. Sustained-release capsules may, depending
on their chemical nature, release the compounds for a few weeks up
to over 100 days. Depending on the chemical nature and the
biological stability of the therapeutic reagent, additional
strategies for protein stabilization may be employed.
The pharmaceutical compositions also may comprise suitable solid-
or gel-phase carriers or excipients. Examples of such carriers or
excipients include calcium carbonate, calcium phosphate, sugars,
starches, cellulose derivatives, gelatin, and polymers such as
polyethylene glycols.
Some of the compounds of the invention may be provided as salts
with pharmaceutically compatible counter ions. Pharmaceutically
compatible salts may be formed with many acids, including
hydrochloric, sulfuric, acetic, lactic, tartaric, malic, succinic,
etc. Salts tend to be more soluble in aqueous or other protonic
solvents than are the corresponding free-base forms.
The agents of the invention may be useful in combination with known
anti-cancer treatments such as: DNA interactive agents such as
cisplatin or doxorubicin; topoisomerase II inhibitors such as
etoposide; topoisomerase I inhibitors such as CPT-11 or topotecan;
tubulin interacting agents such as paclitaxel, docetaxel or the
epothilones; hormonal agents such as tamoxifen; thymidilate
synthase inhibitors such as 5-fluorouracil; and anti-metalbolites
such as methotrexate. They may be administered together or
sequentially, and when administered sequentially, the agents may be
administered either prior to or after administration of the known
anticancer or cytotoxic agent.
The term "chemotherapeutic agent" as used herein includes, for
example, hormonal agents, antimetabolites, DNA interactive agents,
tubilin-interactive agents, and others such as aspariginase or
hydroxyureas.
DNA-interactive agents include alkylating agents, such as
cisplatin, cyclophosphamide, altretamine; DNA strand-breakage
agents, such as bleomycin; intercalating topoisomerase II
inhibitors, e.g., dactinomycin and doxorubicin); nonintercalating
topoisomerase II inhibitors such as, etoposide and teniposide; and
the DNA minor groove binder plicamydin, for example.
Alkylating agents may form covalent chemical adducts with cellular
DNA, RNA, or protein molecules, or with smaller amino acids,
glutathione, or similar chemicals. Examples of typical alkylating
agents include, but are not limited to, nitrogen mustards, such as
chlorambucil, cyclophosphamide, isofamide, mechlorethamine,
melphalan, uracil mustard; aziridine such as thiotepa;
methanesulfonate esters such as busulfan; nitroso ureas, such as
carmustine, lomustine, streptozocin; platinum complexes, such as
cisplatin, carboplatin; bioreductive alkylator, such as mitomycin,
and procarbazine, dacarbazine and altretamine. DNA strand-breaking
agents include bleomycin, for example.
DNA topoisomerase II inhibitors may include intercalators such as
the following: amsacrine, dactinomycin, daunorubicin, doxorubicin
(adriamycin), idarubicin, and mitoxantrone; as well as
nonintercalators such as etoposide and teniposide.
An example of a DNA minor groove binder is plicamycin.
Antimetabolites generally interfere with the production of nucleic
acids and thereby growth of cells by one of two major mechanisms.
Certain drugs inhibit production of deoxyribonucleoside
triphosphates that are the precursors for DNA synthesis, thus
inhibiting DNA replication. Examples of these compounds are
analogues of purines or pyrimidines and are incorporated in
anabolic nucleotide pathways. These analogues are then substituted
into DNA or RNA instead of their normal counterparts.
Antimetabolites useful as chemotherapeutic agents include, but are
not limited to: folate antagonists such as methotrexate and
trimetrexate; pyrimidine antagonists, such as fluorouracil,
fluorodeoxyuridine, CB3717, azacitidine, cytarabine, and
floxuridine; purine antagonists such as mercaptopurine,
6-thioguanine, fludarabine, pentostatin; and ribonucleotide
reductase inhibitors such as hydroxyurea.
Tubulin interactive agents act by binding to specific sites on
tubulin, a protein that polymerizes to form cellular microtubules.
Microtubules are critical cell structure units and are required for
cell division. These therapeutic agents disrupt the formation of
microtubules. Exemplary tubulin-interactive agents include
vincristine and vinblastine, both alkaloids and paclitaxel
(Taxol).
Hormonal agents are also useful in the treatment of cancers and
tumors, but only rarely in the case of B cell malignancies. They
are used in hormonally susceptible tumors and are usually derived
from natural sources. Hormonal agents include, but are not limited
to, estrogens, conjugated estrogens and ethinyl estradiol and
diethylstilbesterol, chlortrianisen and idenestrol; progestins such
as hydroxyprogesterone caproate, medroxyprogesterone, and
megestrol; and androgens such as testosterone, testosterone
propionate; fluoxymesterone, and methyltestosterone.
Adrenal corticosteroids are derived from natural adrenal cortisol
or hydrocortisone and are used to treat B cell malignancies. They
are used because of their anti-inflammatory benefits as well as the
ability of some to inhibit mitotic divisions and to halt DNA
synthesis. These compounds include, but are not limited to,
prednisone, dexamethasone, methylprednisolone, and
prednisolone.
Leutinizing hormone releasing hormone agents or
gonadotropin-releasing hormone antagonists are used primarily the
treatment of prostate cancer. These include leuprolide acetate and
goserelin acetate. They prevent the biosynthesis of steroids in the
testes.
Antihormonal antigens include, for example, antiestrogenic agents
such as tamoxifen, antiandrogen agents such as flutamide; and
antiadrenal agents such as mitotane and aminoglutethimide.
Other agents include hydroxyurea (which appears to act primarily
through inhibition of the enzyme ribonucleotide reductase), and
asparaginase (an enzyme which converts asparagine to aspartic acid
and thus inhibits protein synthesis).
Included within the scope of cancer therapy agents are radiolabeled
antibodies, including but not limited to, Zevalin.TM. (IDEC
Pharmaceuticals Corp.) and Bexxar.TM. (Corixa, Inc.); the use of
any other radioisotope (e.g., .sup.90Y and .sup.131I) coupled to an
antibody or antibody fragment that recognizes an antigen expressed
by a neoplasm; external beam radiation or any other method for
administration of radiation to a patient.
Further included within the scope of cancer therapy agents are
cytotoxins, including but not limited to an antibody or antibody
fragment linked to a cytotoxin, or any other method for selectivly
delivering a cytotoxic agent to a tumor cell.
Further included within the scope of cancer therapy agents are
selective methods for destroying DNA, or any method for delivering
heat to a tumor cells, including by way of example only,
nanoparticles.
Further included within the scope of cancer therapy agents is the
use of unlabeled antibodies or antibody fragments capable of
killing or depleting tumor cells, including by way of example only,
Rituxan.TM. (IDEC Pharmaceuticals Corp.) and Herceptin.TM.
(Genentech).
The agents may be prepared using the reaction routes and synthesis
schemes as described below, employing the general techniques known
in the art using starting materials that are readily available. The
preparation of preferred compounds of the present invention is
described in detail in the following examples, but the artisan will
recognize that the chemical reactions described may be readily
adapted to prepare a number of other anti-proliferatives or protein
kinase inhibitors of the invention. For example, the synthesis of
non-exemplified compounds according to the invention may be
successfully performed by modifications apparent to those skilled
in the art, e.g., by appropriately protecting interfering groups,
by changing to other suitable reagents known in the art, or by
making routine modifications of reaction conditions. Alternatively,
other reactions disclosed herein or generally known in the art will
be recognized as having applicability for preparing other compounds
of the invention.
DETAILED DESCRIPTION
The compounds of Formula (I) can act as antagonists of the VEGFR2.
Without being bound to any particular theory, the linked rings are
thought to provide favorable space-filling and electrostatic
complementarity in the active site of the targeted protein.
In the examples described below, unless otherwise indicated, all
temperatures are set forth in degrees Celsius and all parts and
percentages are by weight. Reagents were purchased from commercial
suppliers such as Aldrich Chemical Company or Lancaster Synthesis
Ltd. and were used without further purification unless otherwise
indicated. Tetrahydrofuran (THF), N,N-dimethylformamide (DMF),
dichloromethane, toluene, and dioxane were purchased from Aldrich
in Sure seal bottles and used as received. All solvents were
purified using standard methods readily known to those skilled in
the art, unless otherwise indicated.
The reactions set forth below were done generally under a positive
pressure of argon or nitrogen or with a drying tube, at ambient
temperature (unless otherwise stated), in anhydrous solvents, and
the reaction flasks were fitted with rubber septa for the
introduction of substrates and reagents via syringe. Glassware was
oven dried and/or heat dried. Analytical thin layer chromatography
(TLC) was performed on glass-backed silica gel 60 F 254 plates
Analtech (0.25 mm) and eluted with the appropriate solvent ratios
(v/v), and are denoted where appropriate. The reactions were
assayed by TLC and terminated as judged by the consumption of
starting material.
Visualization of the TLC plates was done with a p-anisaldehyde
spray reagent or phosphomolybdic acid reagent (Aldrich Chemical 20
wt % in ethanol) and activated with heat. Work-ups were typically
done by doubling the reaction volume with the reaction solvent or
extraction solvent and then washing with the indicated aqueous
solutions using 25% by volume of the extraction volume unless
otherwise indicated. Product solutions were dried over anhydrous
Na.sub.2SO.sub.4 prior to filtration and evaporation of the
solvents under reduced pressure on a rotary evaporator and noted as
solvents removed in vacuo. Flash column chromatography (Still et
al., J. Org. Chem., 43, 2923 (1978)) was done using Baker grade
flash silica gel (47 61 .mu.m) and a silica gel: crude material
ratio of about 20:1 to 50:1 unless otherwise stated. Hydrogenolysis
was done at the pressure indicated in the examples or at ambient
pressure.
.sup.1H-NMR spectra were recorded on a Bruker instrument operating
at 300 MHz and .sup.13C-NMR spectra were recorded operating at 75
MHz. NMR spectra were obtained as CDCl.sub.3 solutions (reported in
ppm), using chloroform as the reference standard (7.25 ppm and
77.00 ppm) or CD.sub.3OD (3.4 and 4.8 ppm and 49.3 ppm), or
internally tetramethylsilane (0.00 ppm) when appropriate. Other NMR
solvents were used as needed. When peak multiplicities are
reported, the following abbreviations are used: s (singlet), d
(doublet), t (triplet), m (multiplet), br (broadened), dd (doublet
of doublets), dt (doublet of triplets). Coupling constants, when
given, are reported in Hertz (Hz).
Infrared (IR) spectra were recorded on a Perkin-Elmer FT-IR
Spectrometer as neat oils, as KBr pellets, or as CDCl.sub.3
solutions, and when given are reported in wave numbers (cm.sup.-1).
The mass spectra were obtained using LSIMS or electrospray. All
melting points (mp) are uncorrected.
General Synthetic Schemes Used for the Preparation of Thenopyridine
Compounds
Preparative Methods
The following methods describe typical synthetic procedures using
specific materials. Many embodiments of the present invention may
be synthesized using the described methods. The skilled artisan
will recognize that different acids, acid chlorides, amines,
phenols, chloropyridine derivatives, and methyl ethers may be
substituted in the following descriptions to suit the preparation
of a desired embodiment. The following methods may be scaled
upwards or downwards to suit the amount of desired material.
##STR00018## (A) Method A Method A follows the general procedure
provided in Scheme I. Scheme I is a general method for amide bond
formation beginning with carboxylic acids and amines. The skilled
artisan will recognize that many methods exist for the coupling of
amines and carboxylates and the method described herein is given by
way of example. (B) Example of Method A To a suspension or solution
of acid, for example
{3-fluoro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]--
phenyl}-acetic acid (126 mg, 0.329 mmol) in CH.sub.2Cl.sub.2 (5 mL)
was added 2 M oxalyl chloride in CH.sub.2Cl.sub.2 (0.49 mL, 0.987
mmol, 3 eq), followed by 3 drops of DMF. The mixture was stirred at
ambient temperature for one hour, concentrated and dried under
vacuum. The crude phenylacetylchloride was re-dissolved in
CH.sub.2Cl.sub.2 (5 mL), and the corresponding amine, for example
2-amino-4,6-dimethylpyridine (60 mg, 0.492 mmol, 1.5 eq) was added,
followed by DMAP (catalytic amount) and triethylamine (1 1.5 eq).
The mixture was stirred at ambient temperature overnight,
concentrated and purified by reversed phase HPLC eluted with 30%
70% acetonitrile in water, or by normal phase silica gel column
eluting with 1% 10% MeOH in CHCl.sub.3 or gradient of EtOAc in
hexanes (depending on the polarity of the product) to give the
desired amide in 20 90% yield.
##STR00019## (A) Method B Method B follows the general procedure
provided in Scheme II. Method B is similar to Method A, but
demonstrates that amide bond formation may precede formation of
thienopyridine-aryl (phenyl) ether bond formation. (B) Example of
Method B To a solution of acid chloride, for example
4-methoxyphenylacetyl chloride (1.00 g, 5.42 mmol) in
CH.sub.2Cl.sub.2 (30 mL) was added amine, such as
2-amino-4,6-dimethylpyridine (661 mg, 5.42 mmol), followed by DMAP
(catalytic amount) and triethylamine (1 eq). After stirring at
ambient temperature overnight, the mixture was concentrated and
purified by flash column chromatography and eluted with gradient of
EtOAc in hexanes to provide the desired amide in 50 90% yield.
##STR00020## (A) Method C Method C follows the general procedure
provided in Scheme III. Method C is a general method for coupling
thienopyridine moieties to phenyl acetic moieties via an ether
linkage. In this method, chloride is displace by phenolate to yield
an aryl phenyl ether. (B) Example of Method C To a solution of
chloropyridine derivative, for example
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine (300
mg, 1.20 mmol), and phenol, for example
3-fluoro-4-hydroxyphenylactic acid (245 mg, 1.44 mmol, 1.2 eq), in
3 ml DMSO was added Cs.sub.2CO.sub.3 (984 mg, 3.00 mmol, 2.5 eq).
The mixture was heated at 100.degree. C. for 12 hours and cooled to
room temperature. EtOAc and water were added, and the mixture was
neutralized by 1N HCl. Precipitate was formed, filtered, and washed
by water. The solid was dried in a vacuum-oven at 60.degree. C. and
used as is for the next step or was further purified by column
chromatography eluting with 1 10% MeOH in CHCl.sub.3. The desired
phenyl ethers were obtained in 40 80% yield.
##STR00021## (A) Method D Method D follows the general procedure
provided in Scheme IV. Method D is a general method for
dealkylating alkyl phenyl ethers to form phenols. (B) Example of
Method D To a 0.degree. C. solution of methyl ether, such as
N-(4,6-dimethyl-pyridin-2-yl)-2-(4-methoxy-phenyl)-acetamide (720
mg, 2.67 mmol), in 15 mL of CH.sub.2Cl.sub.2 was added 1.0 M
BBr.sub.3 (8.00 mL, 8.00 mmol, 3 4 eq). The mixture was stirred at
room temperature overnight. The reaction was quenched with MeOH,
neutralized with concentrated aqueous NH.sub.4OH to pH .about.7.
The resulting mixture was stirred at room temperature for one hour
and poured into water, extracted with CH.sub.2Cl.sub.2 for three
times, dried over Na.sub.2SO.sub.4, concentrated in vacuo, and
further purified by column chromatography to give the desired
phenol or alcohol in 70 100% yield.
EXAMPLES
Example 1
N-(4,6-Dimethyl-pyridin-2-yl)-2-{3-fluoro-4-[2-(1-methyl-1H-imidazol-2-yl)-
-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide
##STR00022##
Intermediate 1a:
{3-Fluoro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]--
phenyl}-acetic acid
##STR00023## was prepared from
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine and
3-fluoro-4-hydroxyphenylactic acid following Method C. The
synthesis of
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine has
been described in PCT application WO99/24440, Example 150. .sup.1H
NMR (300 MHz, DMSO-d.sub.6) .delta. 8.54 (d, 1H, J=4.90 Hz), 7.91
(s, 1H), 7.49 7.42 (m, 3H), 7.26 (d, 1H, J=9.42 Hz), 7.05(s, 1H),
6.65 (d, 1H, J=5.27 Hz), 4.00 (s, 2H), 3.17 (s, 3H). LCMS (ESI+)
[M+H]/z Calc'd 384, found 384.
The compound of Example 1 was prepared from intermediate 1a and
2-amino-4,6-dimethylpyridine following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.65 Hz), 7.73 (s, 1H),
7.65 (s, 1H), 7.35 7.24 (m, 4H), 7.03 (s, 1H), 6.77 (s, 1H), 6.62
(d, 2H, J=4.52 Hz), 3.95 (s, 3H), 3.74 (s, 2H), 2.32 (s, 3H), 2.23
(s, 3H). LCMS (ESI+) [M+H]/z Calc'd 488, found 488. Anal.
(C.sub.26H.sub.22N.sub.5O.sub.2SF.1.0H.sub.2O.1.2CH.sub.3COOH)C, H,
N.
Example 2
2-{3-Fluoro-4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyrid-
in-7-yloxy]-phenyl}-N-methyl-acetamide
##STR00024##
Intermediate 2a:
2-(3-Fluoro-4-hydroxy-phenyl)-N-methyl-acetamide
##STR00025## was prepared from 3-fluoro-4-hydroxyphenylactic acid
and methyl amine following Method A. S.sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.89 (s, 1H), 6.90 (d, 1H, J=13.37 Hz), 6.82
6.71 (m, 1H), 3.29 (s, 2H), 2.62 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 184, found 184.
Intermediate 2b:
(7-Chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e
##STR00026## was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid lithium salt
(prepared according to PCT application WO01/94353, Example 1) and
3R-hydroxy-pyrrolidine following Method A. .sup.1H NMR
(DMSO-d.sub.6): .delta. 8.73 (1H, d, J=5.1 Hz), 8.15, 8.09 (1H, s),
7.69 (1H, d, J=5.1 Hz), 5.10 5.06 (1H, m), 4,43 4.29 (1H, m), 4.05
3.89 (2H, m), 3.72 3.43 (2H, m), 2.08 1.79 (2H, m).
The compound of Example 2 was prepared from the coupling of
intermediates 2a and 2b following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 7.85 (d, 1H, J=17.33
Hz), 7.28 7.12 (m, 3H), 6.62 (d, 1H, J=5.46 Hz), 4.41(bs, 1H), 3.95
3.89 (m, 2H), 3.73 3.60 (m, 3H), 3.47 (s, 2H), 2.65 (s, 3H), 2.13
1.94 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 430, found 430. Anal.
(C.sub.21H.sub.20N.sub.3O.sub.4SF.0.4CH.sub.2Cl.sub.2) C, H, N.
Example 3
2-{4-[2-((3R, 4R)-3,4-Dihydroxy-pyrrolidine-1-carbonyl)-thieno
3,2-b]pyridin-7-yloxy]-phenyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide
##STR00027##
Intermediate 3a:
N-(4,6-Dimethyl-pyridin-2-yl)-2-(4-methoxy-phenyl)-acetamide
##STR00028## was prepared from 4-methoxyphenylacetyl chloride and
2-amino-4,6-dimethylpyridine following Method B. .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 7.86 (s, 1H), 7.73 (s, 1H), 7.24 (d, 2H,
J=8.34 Hz), 6.91 (d, 2H, J=8.59 Hz), 6.70(s, 1H), 3.81 (s, 3H),
3.66 (s, 2H), 2.35 (s, 3H), 2.29 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 271, found 271.
Intermediate 3b:
N-(4,6-Dimethyl-pyridin-2-yl)-2-(4-hydroxy-phenyl)-acetamide
##STR00029## was prepared from intermediate 3a following Method D.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.99 (s, 1H), 7.95 (s,
1H), 7.06 (d, 2H, J=8.34 Hz), 6.74 (s, 1H), 6.63 (d, 2H, J=8.59
Hz), 3.66 (s, 2H), 2.34 (s, 3H), 2.33 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 257, found 257.
Intermediate 3c: 3,4-cis-Dihydroxy-pyrrolidine-1-carboxylic acid
benzyl ester
##STR00030## To a solution of benzyl 3-pyrroline-1-carboxylate (15
g, 90%, 66.4 mmol) in THF (100 mL) and water (25 mL), was added
osmium tetroxide (10 mL, 2.5 wt. % solution in 2-methyl-2-propanol,
0.8 mmol) and 4-methylmorpholine N-oxide (8.56 g, 73 mmol) as
solid. The mixture was stirred at room temperature overnight and
concentrated, in vacuo. The residue was re-dissolved in EtOAc (300
mL) and washed with aqueous Na.sub.2SO.sub.3 (1.5 g in 100 mL
water) solution, aqueous NaHCO.sub.3 solution and brine. The
combined aqueous layer was extracted once with EtOAc (100 mL). The
combined organic extracts were dried over Na.sub.2SO.sub.4 and
concentrated, in vacuo. The crude product was further purified by
flash column chromatography eluting with 4 5% MeOH in
CH.sub.2Cl.sub.2 to give 15.26 g (97%) of a white solid. .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 7.34 (5H, m), 5.11 (2H, bs), 4.26
(2H, m), 3.66 (2H, m), 3.41 (2H, m), 1.56 (2H, bs).
Intermediate 3d: 3,4-cis-Dimethoxy-pyrrolidine-1-carboxylic acid
benzyl ester
##STR00031##
To a stirred solution of 3,4-cis-dihydroxy-pyrrolidine-1-carboxylic
acid benzyl ester 3c (15.2 g, 64.3 mmol) in anhydrous THF (130 mL)
was added iodomethane (36 g, 257 mmol) at 0.degree. C.; sodium
hydride (6.4 g, 60% in mineral oil, 160 mmol) was then added slowly
as at 0.degree. C. The mixture was allowed to warm to room
temperature and stirred at room temperature for 3 hours. Aqueous 1N
HCl (30 mL) was then added to the mixture which was concentrated,
in vacuo, to remove THF. The residue was re-dissolved in EtOAc (300
mL) and washed with water and brine. The organic layer was dried
over Na.sub.2SO.sub.4, filtered, and concentrated, in vacuo. The
crude was further purified by flash column chromatography eluting
with 5 25% EtOAc in CH.sub.2Cl.sub.2, to give 17 g (99%) of
intermediate 3d as a yellow oil. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.35 (5H, m), 5.12 (2H, m), 3.87 (2H, m), 3.55 (2H, m),
3.42 (6H, bs), 1.58 (2H, s).
Intermediate 3e: 3,4-cis-Dimethoxy-pyrrolidine
##STR00032##
To a stirred solution of 3,4-cis-dimethoxy-pyrrolidine-1-carboxylic
acid benzyl ester 3d (16.95 g, 63.9 mmol) in MeOH (150 mL), was
added 10% Pd on C (1.3 g). The mixture was stirred under an H.sub.2
balloon at room temperature for 3 hours and filtered through
celite. The filtrate was concentrated, in vacuo, re-dissolved in
CH.sub.2Cl.sub.2 and dried over Na.sub.2SO.sub.4. The solution was
concentrated to give 8.3 g (99%) of intermediate 3e as a yellow
oil. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 3.80 (2H, m), 3.47
(2H, bs), 3.41 (6H, s), 3.01 (2H, bs).
Intermediate 3f:
7-Chloro-2-[meso-3,4-dimethoxypyrrolidine-1-carbonyl]thieno[3,2-b]pyridin-
e
##STR00033## was prepared by the coupling of lithium
7-chlorothieno[3,2-b]pyridine-2-carboxylate and
3,4-cis-dimethoxypyrrolidine 3e in a manner as previously described
in Method A to give intermediate 3f as a pale yellow syrup. .sup.1H
NMR (CD.sub.3OD): .delta. 8.70 (1H, d, J=5.1 Hz), 8.03 (1H, s),
7.61 (1H, d, J=5.1 Hz), 4.20 4.07 (2H, m), 3.97 3.75 (2H, m), 3.52
(3H, s), 3.48 (3H, s), 3.35 3.29 (2H, m).
Intermediate 3g:
7-Chloro-2-[meso-3,4-dimethoxypyrrolidine-1-carbonyl]thieno[3,2-b]pyridin-
e
##STR00034## was prepared from
7-chloro-2-[meso-3,4-dimethoxypyrrolidine-1-carbonyl]thieno[3,2-b]pyridin-
e (3f) and BBr.sub.3 in a manner described in Method D and gave the
intermediate 3g as a pale white solid.
The compound of Example 3 was prepared from
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3,4-dihydroxy-pyrrolidin-1-yl)-meth-
anone (3g) and
N-(4,6-dimethyl-pyridin-2-yl)-2-(4-hydroxy-phenyl)-acetamide (3b)
following Method C. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.43
(d, 1H, J=5.47 Hz), 7.81 (d, 1H, J=6.60 Hz), 7.64 (s, 1H), 7.42 (d,
2H, J=8.67 Hz), 7.14 (d, 2H, J=8.47 Hz), 6.75 (s, 1H), 6.66 (d, 1H,
J=5.46 Hz), 4.19 (s, 2H), 4.03 3.98 (m, 1H), 3.74 3.68 (m, 4H),
3.57 3.52 (m, 1H), 2.30 (s, 3H), 2.21 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 519, found 519. Anal.
(C.sub.27H.sub.26N.sub.4O.sub.5S.0.6EtOAc.0.2CHCl.sub.3) C, H,
N.
Example 4
2-{4-[2-((R)-3-Dimethylamino-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin--
7-yloxy]-phenyl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide
##STR00035##
Intermediate 4a:
(7-Chloro-thieno[3,2-b]pyridin-2-yl)-(3R-dimethylamino-pyrrolidin-1-yl)-M-
ethanone
##STR00036## was prepared by the reaction of
7-chlorothieno[3,2-b]pyridine-2-carboxylic acid (0.214 g, 1.0 mmol)
with (3R)-N,N-dimethylpyrrolidin-3-amine (0.114 g, 1.0 mmol) and
Et.sub.3N (0.139 mL, 1.0 mmol) in the manner of Method A and gave
intermediate 4a as a brown solid (0.1349, 43%). .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 7.24 (d, 1H, J=5.09 Hz), 6.57 (d, 1H,
J=8.48 Hz), 6.15 (d, 1H, J=5.09 Hz), 2.70 (m, 1H), 2.51 (m, 2H),
2.24 (m, 1H), 2.04 (m, 1H), 1.49 (m, 1H), 0.93 (s, 3H), 0.90 (s,
3H), 0.52 (m, 1H); ESIMS (MH.sup.+): 310.10.
The compound of Example 4 was prepared from
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-dimethylamino-pyrrolidin-1-yl)-me-
thanone (4a) and intermediate 3b following Method C. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 7.84 (d, 1H,
J=6.40 Hz), 7.65 (s, 1H), 7.43 (d, 2H, J=8.48 Hz), 7.16 (d, 2H,
J=8.47 Hz), 6.76 (s, 1H), 6.67 (d, 1H, J=5.46 Hz), 4.14 3.95 (m,
3H), 3.95 3.77 (m, 2H), 3.72 (s, 2H), 3.65 3.35 (m, 2H), 2.31 (s,
6H), 2.22 (s, 6H). LCMS (ESI+) [M+H]/z Calc'd 530, found 530. Anal.
(C.sub.29H.sub.31N.sub.5O.sub.3S.1.0CH.sub.3COOH)C, H, N.
Example 5
7-{4-[(4,6-Dimethyl-pyridin-2-ylcarbamoyl)-methyl]-phenoxy}-thieno[3,2-b]p-
yridine-2-carboxylic acid (3-dimethylamino-propyl)-methyl-amide
##STR00037##
Intermediate 5a: 7-Chloro-thieno[3,2-b]pyridine-2-carboxylic acid
(3-dimethylamino-propyl)-methyl-amide
##STR00038## was prepared by the reaction of
7-chlorothieno[3,2-b]pyridine-2-carbonic acid (1.0 g, 4.68 mmol)
with N,N,N'-trimethylpropane-1,3-diamine (0.868 mL, 4.68 mmol) and
Et.sub.3N (1.96 mL, 14.04 mmol) in a manner as described previously
in Method A and gave intermediate 5a as a white foam (1.07 g, 77%).
.sup.1H NMR (300 MHz, CD.sub.3OD) 38.56 (d, 1H, J=5.09 Hz), 7.76
(s, 1H), 7.46 (d, 1H, J=5.27 Hz), 3.51 (m, 2H), 3.20 (s, 3H), 2.33
(m, 2H), 2.18 (s, 6H), 1.79 (m, 2H); ESIMS (MH.sup.+): 312.05.
The compound of Example 5 was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid
(3-dimethylamino-propyl)-methyl-amide (5a) and intermediate 3b
following Method C. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.42
(d, 1H, J=5.27 Hz), 7.76 (d, 1H, J=8.67 Hz), 7.65 (s, 1H), 7.43 (d,
2H, J=8.48 Hz), 7.15 (d, 2H, J=8.48 Hz), 6.75 (s, 1H), 6.65 (d, 1H,
J=5.65 Hz), 3.71 (s, 2H), 3.56 3.51 (m, 2H), 3.08 (s, 3H), 2.60
2.43 (m, 2H), 2.30 (s, 6H), 2.22 (s, 6H), 1.90 1.81 (m, 2H). LCMS
(ESI+) [M+H]/z Calc'd 532, found 532. Anal.
(C.sub.29H.sub.33N.sub.5O.sub.3S.1.1CH.sub.3COOH)C, H, N.
Example 6
7-{4-[(4,6-Dimethyl-pyridin-2-ylcarbamoyl)-methyl]-phenoxy}-thieno[3,2-b]p-
yridine-2-carboxylic acid (2-dimethylamino-ethyl)-methyl-amide
##STR00039##
Intermediate 6a: 7-Chloro-thieno[3,2-b]pyridine-2-carboxylic acid
(2-dimethylamino-ethyl)-methyl-amide
##STR00040## was prepared by the reaction of
7-chlorothieno[3,2-b]pyridine-2-carboxylic acid (0.957 g, 4.48
mmol) with N,N,N'-trimethylethane-1,2-diamine (0.640 mL, 4.93 mmol)
and Et.sub.3N (0.624 mL, 4.48 mmol) in a manner as described
previously in Method A to give a brown solid (0.167 g, 13%).
.sup.1H NMR (400 MHz, CDCl.sub.3) .delta. 8.60 (d, 1H, J=5.05 Hz),
7.74 (s, 1H), 7.32 (d, 1H, J=5.05 Hz), 3.66 (t, 2H, J=6.19 Hz),
3.26 (s, 3H), 2.57 (t, 2H, J=6.69 Hz), 2.25 (s, 6H). ESIMS
(MH.sup.+): 298.05.
The compound of Example 6 was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid
(2-dimethylamino-ethyl)-methyl-amide (6a) and intermediate 3b
following Method C. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.42
(d, 1H, J=5.56 Hz), 7.75 (m, 1H), 7.65 (s, 1H), 7.44 (d, 2H, J=8.59
Hz), 7.15 (d, 2H, J=8.59 Hz), 6.76 (s, 1H), 6.67 (d, 1H, J=5.56
Hz), 3.72 (s, 2H), 3.22 (s, 5H), 2.86 2.72 (m, 2H), 2.44 (s, 6H),
2.31 (s, 3H), 2.22 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 518, found
518. Anal.
(C.sub.28H.sub.31N.sub.5O.sub.3S.1.0H.sub.2O.0.8CH.sub.3COOH)C, H,
N.
Example 7
7-{4-[(4,6-Dimethyl-pyridin-2-ylcarbamoyl)-methyl]-phenoxy}-thieno[3,2-b]p-
yridine-2-carboxylic acid dimethylamide
##STR00041##
Intermediate 7a: 7-Chloro-thieno[3,2-b]pyridine-2-carboxylic acid
dimethylamide
##STR00042## was prepared by the reaction of
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid (0.57 g, 2.67
mmol) with 2.0 M N,N-dimethylamine in THF (1.60 mL, 3.20 mmol) and
Et.sub.3N (0.447 mL, 3.20 mmol) in a manner as described in Method
A to give the desired amide as brown solid (0.54 g, 84%). .sup.1H
NMR (300 MHz, CDCl.sub.3) .delta. 8.63 (d, 1H, J=4.85 Hz), 7.74 (s,
1H), 7.35 (d, 1H, J=5.02 Hz), 3.28 (s, 3H), 3.22 (s, 3H); ESIMS
(MH.sup.+): 240.95.
The compound of Example 7 was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid dimethylamide (7a)
and intermediate 3b following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.48 (d, 1H, J=5.27 Hz), 7.78 (d, 1H, J=8.67
Hz), 7.60 (s, 1H), 7.48 (d, 2H, J=8.48 Hz), 7.18 (d, 2H, J=8.48
Hz), 6.78 (s, 1H), 6.68 (d, 1H, J=5.65 Hz), 3.78 (s, 2H), 3.14 (s,
3H), 3.08 (s, 3H), 2.33 (s, 3H), 2.28 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 461, found 461. Anal.
(C.sub.25H.sub.24N.sub.4O.sub.3S.1.0H.sub.2O.0.4CH.sub.3COOH)C, H,
N.
Example 8
7-{4-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-phenoxy}-thieno[3,2-b]pyrid-
ine-2-carboxylic acid dimethylamide
##STR00043##
Intermediate 8a:
[4-(2-Dimethylcarbamoyl-thieno[3,2-b]pyridin-7-yloxy)-phenyl]-acetic
acid
##STR00044## was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid dimethylamide (7a)
and 4-hydroxyphenylacetic acid following Method C. .sup.1H NMR (300
MHz, DMSO-d.sub.6) .delta. 8.56 (d, 1H, J=5.31 Hz), 7.92 (s, 1H),
7.40 (d, 2H, J=8.58 Hz), 7.24 (d, 2H, J=8.09 Hz), 6.70 (d, 1H,
J=5.06 Hz), 3.64 (s, 2H), 3.33 (s, 6H). LCMS (ESI+) [M+H]/z Calc'd
357, found 357.
The compound of Example 8 was prepared from intermediate 8a and
2-amino-5-chloro pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.68 (d, 1H, J=5.27 Hz), 8.32 (s, 1H), 8.22 (m,
1H), 7.68 (d, 2H, J=6.03 Hz), 7.43 (d, 2H, J=8.29 Hz), 7.15 (d, 2H,
J=8.48 Hz), 6.70 (d, 1H, J=4.71 Hz), 3.74 (s, 2H), 3.21 (s, 6H).
LCMS (ESI+) [M+H]/z Calc'd 467, found 467. Anal.
(C.sub.23H.sub.19N.sub.4O.sub.3SCl.0.6H.sub.2O.0.6CH.sub.3COOH)C,
H, N.
Example 9
N-(4,6-Dimethyl-pyridin-2-yl)-2-{4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3-
,2-b]pyridin-7-yloxy]-phenyl}-acetamide
##STR00045##
Intermediate 9a:
{4-[2-(1-Methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-a-
cetic acid
##STR00046## was prepared from
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine and
4-hydroxyphenylactic acid following Method C. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 8.51 (d, 1H, J=5.09), 7.88 (s, 1H), 7.40 (m,
3H), 7.24 (d, 2H, J=8.48 Hz), 7.04 (s, 1H), 6.65 (d, 1H, J=5.09
Hz), 3.98 (s, 3H), 3.64 (s, 2H). LCMS (ESI+) [M+H]/z Calc'd 366,
found 366.
The compound of Example 9 was prepared from intermediate 9a and
2-amino-4,6-dimethyl-pyridine following Method A. .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.45 (d, 1H, J=5.84 Hz), 8.19 (d, 1H,
J=5.65 Hz), 7.85 (s, 1H), 7.52 (m, 2H), 7.21 (m, 3H), 7.04(s, 1H),
6.89 (s, 1H), 6.71 (d, 1H, J=6.05 Hz), 3.98 (s, 3H), 3.90 (s, 2H),
2.59 (s, 3H), 2.45 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 470, found
470. Anal.
(C.sub.26H.sub.23N.sub.5O.sub.2S.0.6H.sub.2O.1.0CH.sub.3COOH)C, H,
N.
Example 10
N-(5-Chloro-pyridin-2-yl)-2-{4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b-
]pyridin-7-yloxy]-phenyl}-acetamide
##STR00047## was prepared from intermediate 9a and 2-amino-5-chloro
pyridine following Method A. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 8.46 (d, 1H, J=6.22 Hz), 8.18 (m, 3H), 8.12 (s, 1H), 7.68
(m, 1H), 7.50 (d, 2H, J=8.67 Hz), 7.23(d, 2H, J=8.67 Hz), 7.10 (s,
1H), 6.78 (d, 1H, J=6.22 Hz), 4.04 (s, 3H), 3.82 (s, 2H). LCMS
(ESI+) [M+H]/z Calc'd 476, found 476. Anal.
(C.sub.24H.sub.18N.sub.5O.sub.2SCl.CH.sub.2Cl.sub.2) C, H, N.
Example 11
2-{4-[2-(1-Methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}--
N-(4-methyl-pyridin-2-yl)-acetamide
##STR00048## was prepared from intermediate 9a and 2-amino-4-methyl
pyridine following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.38 (d, 1H, J=5.65 Hz), 8.05 (d, 1H, J=5.28 Hz), 7.86 (s,
1H), 7.71 (s, 1H), 7.44 (d, 2H, J=8.29 Hz), 7.23(s, 1H), 7.16 (d,
2H, J=8.48 Hz), 7.01 (s, 1H), 6.88 (d, 1H, J=4.70 Hz), 6.63 (d, 1H,
J=5.65 Hz), 3.93 (s, 3H), 3.73 (s, 2H), 2.27 (s, 3H). LCMS (ESI+)
[M+H]/z Calc'd 456, found 456.
Example 12
N-Isoquinolin-3-yl-2-(4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridi-
n-7-yloxy]-phenyl}-acetamide
##STR00049## was prepared from intermediate 9a and 3-amino
isoquinoline following Method A. Spectral and analytical data for
the compound of Example 12: .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.97(s, 1H), 8.39 (d, 2H, J=5.46 Hz), 7.90 (s, 1H), 7.71
(m, 2H), 7.51(m, 1H), 7.46 (m, 3H), 7.20 (m, 3H), 7.01 (s, 1H),
6.64 (d, 1H, J=6.22 Hz), 3.94 (s, 3H), 3.82 (s, 2H). LCMS (ESI+)
[M+H]/z Calc'd 492, found 492.
Example 13
2-{4-[2-(1-Methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}--
N-(5-trifluoromethyl-pyridin-2-yl)-acetamide
##STR00050## was prepared from intermediate 9a and
2-amino-5-trifluoro-methyl pyridine following Method A. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.51 (s, 1H), 8.38 (d, 1H, J=5.65
Hz), 8.24 (d, 1H, J=8.67 Hz), 7.96 (d, 1H, J=8.86 Hz), 7.71 (s,
1H), 7.43 (d, 2H, J=8.48 Hz), 7.23 (s, 1H), 7.16 (d, 2H, J=8.67
Hz), 7.01 (s, 1H), 6.62 (d, 1H, J=5.46 Hz), 3.93 (s, 3H), 3.77 (s,
2H). LCMS (ESI+) [M+H]/z Calc'd 510, found 510.
Example 14
N-(4,6-Dimethyl-pyridin-2-yl)-2-{4-[2-((R)-3-hydroxy-pyrrolidine-1-carbony-
l)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide
##STR00051##
Intermediate 14a:
{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy-
]-phenyl}-acetic acid
##STR00052## was prepared from
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) and 4-hydroxyphenylacetic acid following Method C. .sup.1H
NMR (300 MHz, DMSO-d.sub.6) .delta. 8.36 (d, 1H, J=4.89 Hz), 7.77
(s, 1H), 7.29 (d, 2H, J=8.47 Hz), 7.29 (d, 2H, J=8.66 Hz), 6.59 (d,
1H, J=5.47 Hz), 4.36 (bs, 1H), 3.92 3.83 (m, 2H), 3.53 (s, 2H),
3.71 3.60 (m, 3H), 2.01 1.93(m, 2H). LCMS (ESI+) [M+H]/z Calc'd
399, found 399.
The compound of Example 14 was prepared from intermediate 14a and
2-amino-4,6-dimethylpyridine following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.45 (d, 1H, J=5.84 Hz), 7.82 (s, 1H),
7.65 (s, 1H), 7.44 (d, 2H, J=8.48 Hz), 7.11 (d, 2H, J=8.67 Hz),
6.76 (s, 1H), 6.67 (d, 1H, J=5.65 Hz), 4.51 (bs, 1H), 4.01 3.91 (m,
2H), 3.85 (s, 2H), 3.75 3.72 (m, 3H), 2.31 (s, 3H), 2.22 (s, 3H),
2.15 1.94 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 503, found 503. Anal.
(C.sub.27H.sub.26N.sub.4O.sub.4S.0.8H.sub.2O.0.8CH.sub.3COOH)C, H,
N.
Example 15
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-(4-methyl-pyridin-2-yl)-acetamide
##STR00053## was prepared from intermediate 14a and
2-amino-4-methyl pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.44 (d, 1H, J=5.28 Hz), 8.06 (d, 1H, J=5.09
Hz), 7.90 7.81 (m, 2H), 7.45 (d, 2H, J=8.29 Hz), 7.17 (d, 2H,
J=8.11 Hz), 6.89 (d, 1H, J=5.65 Hz), 6.68 (d, 1H, J=5.27 Hz), 4.43
(bs, 1H), 3.98 3.93 (m, 2H), 3.74 (s, 2H), 3.67 3.61(m, 3H), 2.28
(s, 3H), 2.11 1.92(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 489, found
489. Anal.
(C.sub.26H.sub.24N.sub.4O.sub.4S.1.0H.sub.2O.1.0CH.sub.3COOH)C, H,
N.
Example 16
N-(5-Chloro-pyridin-2-yl)-2-{4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-t-
hieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetamide
##STR00054## was prepared from intermediate 14a and
2-amino-5-chloro pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 8.20 (d, 1H, J=2.07
Hz), 8.05 (d, 1H, J=9.04 Hz), 7.85 (d, 1H, J=17.52 Hz), 7.72 7.66
(m, 1H), 7.44 (d, 2H, J=8.48 Hz), 7.16 (d, 2H, J=8.67 Hz), 6.68 (d,
1H, J=5.66 Hz), 4.52 (bs, 1H), 3.99 3.93 (m, 2H), 3.74 (s, 2H),
3.68 3.60(m, 3H), 2.11 1.92(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 509,
found 509.
Example 17
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-isoquinolin-3-yl-acetamide
##STR00055##
was prepared from intermediate 14a and 3-amino-isoquinoline
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.97
(s, 1H), 8.45 8.36 (m, 2H), 7.91 7.81 (m, 2H), 7.74 (d, 1H, J=8.10
Hz), 7.62 7.57 (m, 1H), 7.51 7.41 (m, 3H), 7.18 (d, 2H, J=8.48 Hz),
6.68 (d, 1H, J=5.65 Hz), 4.44 (bs, 1H), 4.02 3.91 (m, 2H), 3.80 (s,
2H), 3.75 3.60 (m, 3H), 2.10 1.91 (m, 2H). LCMS (ESI+) [M+H]/z
Calc'd 525, found 525. Anal.
(C.sub.29H.sub.24N.sub.4O.sub.4S.0.8CH.sub.2Cl.sub.2) C, H, N.
Example 18
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-(5-trifluoromethyl-pyridin-2-yl)-acetamide
##STR00056## was prepared from intermediate 14a and
2-amino-5-trifluoro-methyl pyridine following Method A. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.51 (s, 1H), 8.42 (d, 1H, J=5.27
Hz), 8.23 (d, 1H, J=8.29 Hz), 7.95 (d, 1H, J=8.48 Hz), 7.84 (d, 1H,
J=17.71 Hz), 7.43 (d, 2H, J=8.29 Hz), 7.15 (d, 2H, J=8.66 Hz), 6.66
(d, 1H, J=5.47 Hz), 4.41 (bs, 1H), 3.99 3.91 (m, 2H), 3.77 (s, 2H),
3.71 3.59 (m, 3H), 2.07 1.98 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd
543, found 543.
Example 19
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-pyridin-2-yl-acetamide
##STR00057## was prepared from intermediate 14a and 2-aminopyridine
following Method A. Spectral and analytical data for the compound
of Example 19: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.53 (d,
1H, J=5.27 Hz), 8.27 (s, 1H), 8.11 (d, 1H, J=8.48 Hz), 7.98 (d, 1H,
J=17.71 Hz), 7.85 7.71 (m, 1H), 7.52 (d, 2H, J=8.66 Hz), 7.31 (d,
2H, J=8.66 Hz), 7.18 7.09 (m, 1H), 6.72 (d, 1H, J=5.47 Hz), 4.42
(bs, 1H), 4.18 3.98 (m, 2H), 3.88 3.55 (m, 5H), 2.21 1.98 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 475, found 475. Anal.
(C.sub.25H.sub.22N.sub.4O.sub.4S.1.2CH.sub.2Cl.sub.2) C, H, N.
Example 20
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-phenyl-acetamide
##STR00058## was prepared from intermediate 14a and aniline
following Method A. Spectral and analytical data for the compound
of Example 20: .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.41 (d,
1H, J=5.65 Hz), 7.83 (d, 1H, J=17.52 Hz), 7.53 7.38 (m, 4H), 7.26
7.11 (m, 4H), 7.03 6.95 (m, 1H), 6.65 (d, 1H, J=5.65 Hz), 4.42 (bs,
1H), 4.04 3.89 (m, 2H), 3.76 3.56 (m, 5H), 2.12 1.98 (m, 2H). LCMS
(ESI+) [M+H]/z Calc'd 474, found 474. Anal.
(C.sub.26H.sub.23N.sub.3O.sub.4S.0.6CH.sub.2Cl.sub.2) C, H, N.
Example 21
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(6--
methoxy-pyridin-3-yl)-acetamide
##STR00059##
Intermediate 21a:
Azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone
##STR00060## was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid and azetidine
hydrochloride following Method A. .sup.1H NMR (300 MHz,
DMSO-d.sub.6) .delta. 8.72 (1H, d, J=5.1 Hz), 7.96 (1H, s), 7.70
(1H, d, J=5.1 Hz), 4.62 (2H, t, J=7.4 Hz), 4.12 (2H, t, J=7.7 Hz),
2.34 (2H, tt, J=7.4, 7.7 Hz).
Intermediate 21b:
{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-acetic
acid
##STR00061## was prepared from
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
and 4-hydroxyphenylacetic acid following Method C. .sup.1H NMR (300
MHz, DMSO-d.sub.6) .delta. 8.57 (d, 1H, J=5.27 Hz), 7.88(s, 1H),
7.40 (d, 2H, J=8.10 Hz), 7.24 (d, 2H, J=8.10 Hz), 6.70 (d, 1H,
J=5.08 Hz), 4.70 4.52 (m, 2H), 4.18 4.00 (m, 2H), 3.64 (s, 2H),
2.46 2.34(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 369, found 369.
The compound of Example 21 was prepared from intermediate 21b and
5-amino-2-methoxy-pyridine following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.66 Hz), 8.23 (d, 1H,
J=2.45 Hz), 7.83 7.79 (m, 1H), 7.74 (s, 1H), 7.43 (d, 2H, J=8.48
Hz), 7.15 (d, 2H, J=8.67 Hz), 6.71 6.66 (m, 2H), 4.64 4.57 (m, 2H),
4.22 4.13 (m, 2H), 3.79 (s, 3H), 3.67 (s, 2H), 2.44 2.34(m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 475, found 475.
Example 22
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(4,-
6-dimethyl-pyridin-2-yl)-acetamide
##STR00062## was prepared from intermediate 21b and
2-amino-4,6-dimethylpyridine following Method A. .sup.1H NMR (300
MHz, CDCl.sub.3) .delta. 8.50 (d, 1H, J=5.27 Hz), 8.03 (s, 1H),
7.77 (s, 1H), 7.47 (d, 2H, J=8.29 Hz), 7.18 (d, 2H, J=8.29 Hz),
6.81 (s, 1H), 6.64 (d, 1H, J=5.28 Hz), 4.67 4.63 (m, 2H), 4.34
4.22(m, 2H), 3.82 (s, 2H), 2.48 (s, 3H), 2.46 2.40(m, 2H), 2.37 (s,
3H). LCMS (ESI+) [M+H]/z Calc'd 473, found 473. Anal.
(C.sub.26H.sub.24N.sub.4O.sub.3S.0.85CH.sub.2Cl.sub.2.0.5EtOAc) C,
H, N.
Example 23
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(4--
methyl-pyridin-2-yl)-acetamide
##STR00063## was prepared from intermediate 21b and
2-amino-4-methyl pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.47 Hz), 8.05 ((d, 1H, J=5.09
Hz), 7.85 (s, 1H), 7.73 (s, 1H), 7.43 (d, 2H, J=8.48 Hz), 7.15 (d,
2H, J=8.66 Hz), 6.88 ((d, 1H, J=5.65 Hz), 6.66 (d, 1H, J=5.65 Hz),
4.65 4.54 (m, 2H), 4.22 4.12(m, 2H), 3.74 (s, 2H), 2.45 2.34(m,
2H), 2.27 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 459, found 459. Anal.
(C.sub.25H.sub.22N.sub.4O.sub.3S.0.4CH.sub.2Cl.sub.2) C, H, N.
Example 24
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
methyl-pyridin-2-yl)-acetamide
##STR00064## was prepared from intermediate 21b and
2-amino-5-methyl pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.46 Hz), 8.05 ((s, 1H), 7.88
(d, 1H, J=8.48 Hz), 7.75 (s, 1H), 7.54 7.49 (m, 1H), 7.43 (d, 2H,
J=8.67 Hz), 7.15 (d, 2H, J=8.66 Hz), 6.66 (d, 1H, J=5.47 Hz), 4.65
4.56 (m, 2H), 4.22 4.12 (m, 2H), 3.73 (s, 2H), 2.45 2.34 (m, 2H),
2.21 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 459, found 459. Anal.
(C.sub.25H.sub.22N.sub.4O.sub.3S.0.3CH.sub.2Cl.sub.2) C, H, N.
Example 25
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(6--
methyl-pyridin-2-yl)-acetamide
##STR00065## was prepared from intermediate 21b and
2-amino-6-methyl pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.65 Hz), 7.78 (d, 1H, J=8.29
Hz), 7.72 (s, 1H), 7.56 7.51 (m, 1H), 7.43 (d, 2H, J=8.66 Hz), 7.15
(d, 2H, J=8.48 Hz), 6.88 (d, 1H, J=7.54 Hz), 6.66 (d, 1H, J=5.47
Hz), 4.60 4.55 (m, 2H), 4.21 4.12 (m, 2H), 3.72 (s, 2H), 2.45 2.36
(m, 2H), 2.34 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 459, found 459.
Anal. (C.sub.25H.sub.22N.sub.4O.sub.3S.0.6EtOAc.0.4H.sub.2O)C, H,
N.
Example 26
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(3--
methyl-pyridin-2-yl)-acetamide
##STR00066## was prepared from intermediate 21b and
2-amino-3-methyl pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.46 Hz), 8.15 (d, 1H, J=4.52
Hz), 7.71 (s, 1H), 7.62 (d, 1H, J=7.72 Hz), 7.45 (d, 2H, J=8.29
Hz), 7.19 7.06 (m, 3H), 6.63 (d, 1H, J=5.47 Hz), 4.63 4.52 (m, 2H),
4.21 4.08 (m, 2H), 3.73 (s, 2H), 2.42 2.29 (m, 2H), 2.11 (s, 3H).
LCMS (ESI+) [M+H]/Z Calc'd 459, found 459.
Example 27
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
trifluoromethyl-pyridin-2-yl)-acetamide
##STR00067## was prepared from intermediate 21b and
2-amino-5-trifluoro-methyl pyridine following Method A. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.61 (s, 1H), 8.42 (d, 1H, J=5.65
Hz), 8.22 (d, 1H, J=8.85 Hz), 7.95 (d, 1H, J=9.04 Hz), 7.74 (s,
1H), 7.43 (d, 2H, J=8.48 Hz), 7.15 (d, 2H, J=8.48 Hz), 6.66 (d, 1H,
J=5.46 Hz), 4.63 4.55 (m, 2H), 4.21 4.12(m, 2H), 3.76 (s, 2H), 2.45
2.36 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 513, found 513.
Example 28
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
chloro-pyridin-2-yl)-acetamide
##STR00068## was prepared from intermediate 21b and
2-amino-5-chloro pyridine following Method A. .sup.1H. NMR (300
MHz, CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.46 Hz), 8.18 (d, 1H,
J=2.07 Hz), 8.03 (d, 1H, J=8.85 Hz), 7.73 (s, 1H), 7.70 7.66 (m,
1H), 7.42 (d, 2H, J=8.48 Hz), 7.15 (d, 2H, J=8.48 Hz), 6.66 (d, 1H,
J=5.46 Hz), 4.63 4.55 (m, 2H), 4.21 4.12 (m, 2H), 3.72 (s, 2H),
2.45 2.36 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 479, found 479. Anal.
(C.sub.24H.sub.19N.sub.4O.sub.3SCl.0.5H.sub.2O.0.8CH.sub.3COOH)C,
H, N.
Example 29
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
fluoro-pyridin-2-yl)-acetamide
##STR00069## was prepared from intermediate 21b and
2-amino-5-fluoro pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.46 Hz), 8.12 (d, 1H, J=3.20
Hz), 8.10 8.02 (m, 1H), 7.75 (s, 1H), 7.64 7.48 (m, 1H), 7.45 (d,
2H, J=8.67 Hz), 7.15 (d, 2H, J=8.48 Hz), 6.67 (d, 1H, J=5.65 Hz),
4.66 4.57 (m, 2H), 4.21 4.12(m, 2H), 3.73 (s, 2H), 2.45 2.36 (m,
2H). LCMS (ESI+) [M+H]/z Calc'd 463, found 463. Anal.
(C.sub.24H.sub.19N.sub.4O.sub.3SF.0.8CH.sub.3COOH)C, H, N.
Example 30
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
bromo-pyridin-2-yl)-acetamide
##STR00070## was prepared from intermediate 21b and 2-amino-5-bromo
pyridine following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.42 (d, 1H, J=5.46 Hz), 8.29 (d, 1H, J=2.64 Hz), 7.99 (d,
1H, J=8.67 Hz), 7.85 7.78 (m, 1H), 7.74 (s, 1H), 7.43 (d, 2H,
J=8.67 Hz), 7.15 (d, 2H, J=8.66 Hz), 6.67 (d, 1H, J=5.46 Hz), 4.66
4.55 (m, 2H), 4.21 4.12 (m, 2H), 3.73 (s, 2H), 2.45 2.36 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 524, found 524. Anal.
(C.sub.24H.sub.19N.sub.4O.sub.3SBr.1.0CH.sub.3COOH)C, H, N.
Example 31
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-iso-
quinolin-3-yl-acetamide
##STR00071## was prepared from intermediate 21b and
3-aminoisoquinoline following Method A. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.95 (s, 1H), 8.61 (s, 1H), 8.52 (d, 1H, J=5.66
Hz), 8.41 (s, 1H), 7.92 7.88 (m, 2H), 7.81 (d, 1H, J=8.29 Hz), 7.70
7.63 (m, 1H), 7.50 (d, 2H, J=8.48 Hz), 7.22 (d, 2H, J=8.66 Hz),
6.73 (d, 1H, J=5.66 Hz), 4.66 4.55 (m, 2H), 4.34 4.24 (m, 2H), 3.86
(s, 2H), 2.52 2.39 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 495, found
495. Anal. (C.sub.28H.sub.22N.sub.4O.sub.3S.0.4CH.sub.2Cl.sub.2) C,
H, N.
Example 32
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-iso-
quinolin-1-yl-acetamide
##STR00072## was prepared from intermediate 21b and
1-aminoisoquinoiine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.56 Hz), 8.20 (d, 1H, J=5.81
Hz), 7.99 7.93 (m, 1H), 7.86 (d, 1H, J=8.08 Hz), 7.73 (s, 1H), 7.73
7.66 (m, 1H), 7.62 (d, 1H, J=5.81 Hz), 7.58 7.54 (m, 1H), 7.51 (d,
2H, J=8.33 Hz), 7.17 (d, 2H, J=8.34 Hz), 6.64 (d, 1H, J=5.56 Hz),
4.63 4.55 (m, 2H), 4.21 4.12 (m, 2H), 3.89 (s, 2H), 2.45 2.33 (m,
2H). LCMS (ESI+) [M+H]/z Calc'd 495, found 495.
Example 33
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-qui-
nolin-2-yl-acetamide
##STR00073## was prepared from intermediate 21b and
2-aminoquinoline following Method A. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.71 (d, 1H, J=9.23 Hz), 8.60 (d, 1H, J=9.42
Hz), 8.54 (d, 1H, J=6.03 Hz), 8.19 (s, 1H), 8.01 7.93 (m, 3H), 7.75
(d, 1H, J=5.84 Hz), 7.68 (d, 2H, J=8.29 Hz), 7.26 (d, 2H, J=8.48
Hz), 7.01 (d, 1H, J=6.22 Hz), 4.71 4.63 (m, 2H), 4.32 4.25 (m, 2H),
4.14 (s, 2H), 2.63 2.43 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 495,
found 495.
Example 34
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-pyr-
idin-3-yl-acetamide
##STR00074## was prepared from intermediate 21b and 3-aminopyridine
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.8.66
(d, 1H, J=2.26 Hz), 8.42 (d, 1H, J=5.46 Hz), 8.19 8.15 (m, 1H),
8.07 8.02 (m, 1H), 7.73 (s, 1H), 7.43 (d, 2H, J=8.48 Hz), 7.34 7.27
(m, 1H), 7.15 (d, 2H, J=8.33 Hz), 6.66 (d, 1H, J=5.47 Hz), 4.63
4.55 (m, 2H), 4.21 4.12 (m, 2H), 3.71 (s, 2H), 2.45 2.33 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 445, found 445. Anal.
(C.sub.24H.sub.20N.sub.4O.sub.3S.0.3CH.sub.2Cl.sub.2.0.4EtOAc) C,
H, N.
Example 35
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-pyr-
idin-4-yl-acetamide
##STR00075## was prepared from intermediate 21b and 4-aminopyridine
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.42
(d, 1H, J=5.46 Hz), 8.30 (d, 2H, J=6.21 Hz), 7.74 (s, 1H), 7.58 (d,
2H, J=6.60 Hz), 7.43 (d, 2H, J=8.67 Hz), 7.15 (d, 2H, J=8.48 Hz),
6.66 (d, 1H, J=5.65 Hz), 4.63 4.55 (m, 2H), 4.21 4.12 (m, 2H), 3.72
(s, 2H), 2.45 2.33 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 445, found
445.
Example 36
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(1H-
-indazol-5-yl)-acetamide
##STR00076## was prepared from intermediate 21b and 5-aminoindazole
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.42
(d, 1H, J=6.32 Hz), 8.00 (bs, 1H), 7.74 (s, 1H), 7.46 (d, 2H,
J=9.36 Hz), 7.40 (s, 1H), 7.80 (d, 2H, J=8.34 Hz), 7.15 (d, 2H,
J=10.11 Hz), 6.66 (d, 1H, J=5.81 Hz), 4.63 4.55 (m, 2H), 4.21 4.12
(m, 2H), 3.70 (s, 2H), 2.43 2.36 (m, 2H). LCMS (ESI+) [M+H]/z
Calc'd 484, found 484.
Example 37
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-ben-
zothiazol-6-yl-acetamide
##STR00077## was prepared from intermediate 21b and
6-aminobenzothiazole following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 9.06 (s, 1H), 8.45 (1H, J=2.02 Hz), 8.42 (d,
1H, J=5.56 Hz), 7.91 (1H, J=8.85 Hz)), 7.74 (s, 1H), 7.54 7.49 (m,
1H), 7.45 (d, 2H, J=8.59 Hz), 7.16 (d, 2H, J=8.59 Hz), 6.67 (d, 1H,
J=5.56 Hz), 4.64 4.53 (m, 2H), 4.21 4.12 (m, 2H), 3.73 (s, 2H),
2.43 2.36 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 501, found 501. Anal.
(C.sub.26H.sub.20N.sub.4O.sub.3S.sub.2.0.2CH.sub.2Cl.sub.2) C, H,
N.
Example 38
2{-4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(6--
morpholin-4-yl-pyridin-3-yl)-acetamide
##STR00078## was prepared from intermediate 21b and
6-morpholin-4-yl-pyridin-3-ylamine following Method A. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 8.33 (d, 1H,
J=2.26 Hz), 7.78 7.72 (m, 2H), 7.43 (d, 2H, J=8.48 Hz), 7.15 (d,
2H, J=8.48 Hz), 6.73 (d, 1H, J=9.04 Hz), 6.67 (d, 1H, J=5.47 Hz),
4.63 4.55 (m, 2H), 4.21 4.15(m, 2H), 3.74 3.64 (m, 6H), 3.37 3.30
(m, 4H), 2.45 2.33 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 530, found
530. Anal.
(C.sub.28H.sub.27N.sub.5O.sub.4S.0.1CH.sub.2Cl.sub.2.1.0MeOH) C, H,
N.
Example 39
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
methyl-1H-pyrazol-3-yl)-acetamide
##STR00079## was prepared from intermediate 21b and
3-amino-5-methylpyrazole following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 7.75 (s, 1H), 7.43 (d,
2H, J=8.48 Hz), 7.15 (d, 2H, J=8.48 Hz), 6.66 (d, 1H, J=5.47 Hz),
5.19 (s, 1H), 4.66 4.57 (m, 2H), 4.21 4.15(m, 2H), 3.68 (s, 2H),
2.45 2.33 (m, 2H), 2.07 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 448,
found 448. Anal. (C.sub.23H.sub.21N.sub.5O.sub.3S.1.1H.sub.2O) C,
H, N.
Example 40
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(1H-
-pyrazol-3-yl)-acetamide
##STR00080## was prepared from intermediate 21b and 2-aminopyrazole
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.43
(d, 1H, J=5.46 Hz), 7.75 (s, 1H), 7.43 (d, 3H, J=8.48 Hz), 7.15 (d,
2H, J=8.48 Hz), 6.67 (d, 1H, J=5.47 Hz), 6.46 6.42 (m, 1H), 4.66
4.57 (m, 2H), 4.21 4.15(m, 2H), 3.68 (s, 2H), 2.45 2.33 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 434, found 434.
Example 41
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(5--
methyl-isoxazol-3-yl)-acetamide
##STR00081## was prepared from intermediate 21b and
3-amino-5-methyl isoxazole following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.65 Hz), 7.75 (s, 1H),
7.40 (d, 2H, J=8.48 Hz), 7.15 (d, 2H, J=8.66 Hz), 6.66 (d, 1H,
J=5.65 Hz), 6.51 (s, 1H), 4.66 4.57 (m, 2H), 4.21 4.15 (m, 2H),
3.70 (s, 2H), 2.45 2.33 (m, 2H), 2.07 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 449, found 449. Anal.
(C.sub.23H.sub.20N.sub.4O.sub.4S.0.55CH.sub.2Cl.sub.2) C, H, N.
Example 42
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-iso-
xazol-3-yl-acetamide
##STR00082## was prepared from intermediate 21b and
3-aminoisoxazole following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.43 (m, 2H), 7.74 (s, 1H), 7.42 (d, 2H, J=8.67
Hz), 7.16 (d, 2H, J=8.48 Hz), 6.66 (d, 1H, J=5.46 Hz), 6.51 (s,
1H), 4.66 4.57 (m, 2H), 4.21 4.15 (m, 2H), 3.72 (s, 2H), 2.45 2.33
(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 435, found 435. Anal.
(C.sub.22H.sub.18N.sub.4O.sub.4S.0.2CH.sub.2Cl.sub.2) C, H, N.
Example 43
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-(3,-
4-dimethyl-isoxazol-5-yl)-acetamide
##STR00083## was prepared from intermediate 21b and
3,4-dimethyl-5-amino isoxazole following Method A.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.44 (d, 1H, J=5.47 Hz),
7.75 (s, 1H), 7.42 (d, 2H, J=8.10 Hz), 7.16 (d, 2H, J=8.48 Hz),
6.67 (d, 1H, J=5.46 Hz), 4.66 4.57 (m, 2H), 4.21 4.15(m, 2H), 3.72
(s, 2H), 2.45 2.33 (m, 2H), 2.11 (s, 3H), 1.77 (s, 3H). LCMS (ESI+)
[M+H]/z Calc'd 463, found 463.
Example 44
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-thi-
azol-2-yl-acetamide
##STR00084## was prepared from intermediate 21b and 2-aminothiazole
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.43
(d, 1H, J=5.56 Hz), 7.74 (s, 1H), 7.42 (d, 2H, J=8.59 Hz), 7.34 (d,
1H, J=5.56 Hz), 7.16 (d, 2H, J=8.59 Hz), 7.03 (d, 1H, J=3.54 Hz),
6.66 (d, 1H, J=5.31 Hz), 4.66 4.57 (m, 2H), 4.21 4.15 (m, 2H), 3.79
(s, 2H), 2.45 2.33 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 451, found
451.
Example 45
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-cyc-
lopropyl-acetamide
##STR00085## was prepared from intermediate 21b and
cyclopropylamine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.46 Hz), 7.73 (s, 1H), 7.34 (d,
2H, J=8.48 Hz), 7.12 (d, 2H, J=8.48 Hz), 6.63 (d, 1H, J=5.47 Hz),
4.66 4.57 (m, 2H), 4.21 4.15 (m, 2H), 3.43 (s, 2H), 2.64 2.56 (m,
1H), 2.45 2.33 (m, 2H), 0.68 0.62 (m, 2H), 0.45 0.39 (m, 2H). LCMS
(ESI+) [M+H]/z Calc'd 408, found 408. Anal.
(C.sub.22H.sub.21N.sub.3O.sub.3S.0.4CH.sub.2Cl.sub.2) C, H, N.
Example 46
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-cyc-
lobutyl-acetamide
##STR00086## was prepared from intermediate 21b and cyclobutylamine
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.42
(d, 1H, J=5.46 Hz), 7.74 (s, 1H), 7.35 (d, 2H, J=8.48 Hz), 7.12 (d,
2H, J=8.48 Hz), 6.64 (d, 1H, J=5.47 Hz), 4.66 4.57 (m, 2H), 4.21
4.15(m, 3H), 3.44 (s, 2H), 2.45 2.33 (m, 2H), 2.25 2.13 (m, 2H),
1.96 1.81 (m, 2H), 1.72 1.69 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd
422, found 422.
Example 47
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-cyc-
lopentyl-acetamide
##STR00087## was prepared from intermediate 21b and
cyclopentylamine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 7.74 (s, 1H), 7.35 (d,
2H, J=8.48 Hz), 7.13 (d, 2H, J=8.66 Hz), 6.64 (d, 1H, J=5.46 Hz),
4.664.57 (m, 2H), 4.21 4.15(m, 2H), 4.07 3.98 (m, 1H), 3.45 (s,
2H), 2.45 2.33 (m, 2H), 1.91 1.81 (m, 2H), 1.69 1.60 (m, 2H), 1.57
1.47 (m, 2H), 1.46 1.34 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 436,
found 436.
Example 48
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-cyc-
lohexyl-acetamide
##STR00088## was prepared from intermediate 21b and cyclohexylamine
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.43
(d, 1H, J=5.47 Hz), 7.74 (s, 1H), 7.35 (d, 2H, J=8.48 Hz), 7.13 (d,
2H, J=8.48 Hz), 6.64 (d, 1H, J=5.47 Hz), 4.66 4.57 (m, 2H), 4.21
4.15 (m, 2H), 3.60 3.52 (m, 1H), 3.45 (s, 2H), 2.45 2.33 (m, 2H),
1.83 1.62 (m, 5H), 1.33 1.06 (m, 5H). LCMS (ESI+) [M+H]/z Calc'd
450, found 450.
Example 49
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(4,6-dimethyl-pyridin-2-yl)-acetamide
##STR00089##
Intermediate 49a: 1-Bromomethyl-2-chloro-4-methoxy-benzene
##STR00090## A suspension of 3-chloro-4-methylanisole (2.23 g,
14.24 mmol), N-bromosuccinimide (2.53 g, 14.24 mmol), and 70%
benzoyl peroxide (493 mg, 1.424 mmol) in 40 mL CCl.sub.4 was heated
to reflux at 80.degree. C. for two hours. The mixture was cooled to
room temperature, filtered, and the filtrate was concentrated under
rotary evaporator. The residue was purified by flash column
chromatography eluting with 5% EtOAc in Hexane to provide 2.25 g of
intermediate 49a as a white solid. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.33 (d, 1H, J=8.48 Hz), 6.93 (d, 1H, J=2.63
Hz), 6.81 6.75 (m, 1H), 4.58 (m, 2H), 3.79 (m, 3H).
Intermediate 49b: (2-Chloro-4-methoxy-phenyl)-acetonitrile
##STR00091## To a solution of
1-bromomethyl-2-chloro-4-methoxy-benzene (49a) (1.67 g, 7.14 mmol)
in methylene chloride (15 mL) was added tetraethylammonium cyanide
(1.67 g, 10.71 mmol). The mixture was stirred at room temperature
for two hours, poured into water, and extracted with EtOAc for
three times. The combined organic layer was dried over MgSO.sub.4,
and concentrated under rotary evaporator. The residue was purified
by flash column chromatography eluting with 10% EtOAc in hexane to
provide 1.13 g of intermediate 49b as a white solid. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 7.37 (d, 1H, J=8.48 Hz), 6.96 (d, 1H,
J=2.64 Hz), 6.86 6.80 (m, 1H), 3.80 (m, 3H), 3.75 (m, 2H).
Intermediate 49c: (2-Chloro-4-methoxy-phenyl)-acetic acid
##STR00092## To a solution of
(2-chloro-4-methoxy-phenyl)-acetonitrile (49b) (1.13 g, 6.24 mmol)
in acetic acid (6 mL) and water (6 mL) was added dropwise
concentrated H.sub.2SO.sub.4 (6 mL). The mixture was refluxed for
eight hours, cooled to room temperature, poured into ice-water,
adjusted pH to .about.9 by aqueous NaOH solution, and washed with
EtOAc. The aqueous layer was acidified with concentrated HCl
aqueous solution to pH 5 and extracted with EtOAc three times, and
combined organic extracts were dried over MgSO.sub.4, concentrated
in vacuo, to give 960 mg of intermediate 49c as a white solid.
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 12.33 (s, 1H), 7.29 (d,
1H, J=8.48 Hz), 7.02 (d, 1H, J=2.64 Hz), 6.90 6.83 (m, 1H), 3.75
(m, 3H), 3.61 (m, 2H). LCMS (ESI-) [M-H]/z Calc'd 199, found
199.
Intermediate 49d: (2-Chloro-4-methoxy-phenyl)-acetic acid methyl
ester
##STR00093## To a solution of (2-chloro-4-methoxy-phenyl)-acetic
acid (49c) (0.86 g, 4.30 mmol) in 20 mL of MeOH was added 0.5 mL of
4.0 M HCl in dioxane. The mixture was stirred at room temperature
overnight, concentrated, poured into water, and extracted three
times with EtOAc. The combined organic layers were dried over
Na.sub.2SO.sub.4, concentrated in vacuo, to give 870 mg of the
intermediate 49d as a colorless syrup. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.18 (d, 1H, J=8.67 Hz), 6.93 (d, 1H, J=2.64
Hz), 6.80 6.75 (m, 1H), 3.78 (m, 3H), 3.70 (m, 2H). LCMS (ESI+)
[M+H]/z Calc'd 215, found 215.
Intermediate 49e: (2-Chloro-4-hydroxy-phenyl)-acetic acid methyl
ester
##STR00094## To a solution of (2-chloro-4-methoxy-phenyl)-acetic
acid methyl ester (49d) (870 mg, 4.06 mmol) in 3 mL of
CH.sub.2Cl.sub.2 was added 1.0 M BBr.sub.3 (12.2 mL, 12.20 mmol),
the mixture was stirred at ambient temperature overnight. The
reaction was quenched with methanol, neutralized with concentrated
aqueous NH.sub.4OH to pH .about.7. The resulting mixture was
stirred at room temperature for one hour, poured into water, and
extracted three times with CH.sub.2Cl.sub.2. The combined organic
layer were dried over Na.sub.2SO.sub.4, concentrated in vacuo to
give 740 mg of the intermediate 49e as a yellow syrup. .sup.1H NMR
(300 MHz, CDCl.sub.3 .delta. 7.09 (d, 1H, J=8.48 Hz), 6.87 (d, 1H,
J=2.63 Hz), 6.69 6.62 (m, 1H), 3.72 (m, 3H), 3.69 (m, 2H). LCMS
(ESI+) [M+H]/z Calc'd 201, found 201.
Intermediate 49f:
{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-pheny-
l}-acetic acid methyl ester
##STR00095## A mixture of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
(930 mg, 3.70 mmol), (2-chloro-4-hydroxy-phenyl)-acetic acid methyl
ester (49e) (740 mg, 3.70 mmol), and Cs.sub.2CO.sub.3 (1.82 g, 7.40
mmol) in 7 mL of DMSO was heated at 100.degree. C. for overnight
and cooled to room temperature. EtOAc and water were added. The
organic layer was washed three times with water, dried over
Na.sub.2SO.sub.4, and concentrated in vacuo. The residue was
purified by flash column chromatography eluting with
EtOAc:CHCl.sub.3:MeOH (1:1:0.04) to provide 640 mg of white solid
as the intermediate 49f. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
8.47 (d, 1H, J=5.46 Hz), 7.74 (s, 1H), 7.43 (d, 1H, J=8.48 Hz),
7.32 (d, 1H, J=2.45 Hz), 7.15 7.10 (m, 1H), 6.73 (d, 1H, J=5.47
Hz), 4.67 4.54 (m, 2H), 4.24 4.13 (m, 2H), 3.80 (s, 3H), 3.65 (s,
2H), 2.48 2.33 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 417, found
417.
Intermediate 49g:
{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-pheny-
l}-acetic acid
##STR00096## To a solution of
{4-[2-(azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-pheny-
l}-acetic acid methyl ester (49f) (0.64 g, 1.54 mmol) in 15 mL of
THF was added 8 mL of 0.33 N KOH at 0.degree. C. The mixture was
stirred at room temperature for three hours, and concentrated in
vacuo. Water was added. The aqueous layer was acidified with 1 N
HCl until precipitate was formed. The solid was filtered, and
washed with water. The solid was dried in a vacuum-oven at
60.degree. C. overnight. The intermediate 49g (600 mg) was obtained
as a white solid. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.47
(d, 1H, J=5.46 Hz), 7.74 (s, 1H), 7.43 (d, 1H, J=8.48 Hz), 7.32 (d,
1H, J=2.45 Hz), 7.15 7.10 (m, 1H), 6.73 (d, 1H, J=5.47 Hz), 4.67
4.54 (m, 2H), 4.24 4.13 (m, 2H), 3.65 (s, 2H), 2.48 2.33 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 403, found 403.
The compound of Example 49 was prepared from intermediate 49g and
2-amino-4,6-dimethyl pyridine following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.47 (d, 1H, J=5.47 Hz), 7.75 (s, 1H),
7.65 (s, 1H), 7.47 (d, 2H, J=8.29 Hz), 7.33 (d, 1H, J=2.45 Hz),
7.18 7.11 (m, 1H), 6.69 6.75 (m, 1H), 4.67 4.63 (m, 2H), 4.34 4.22
(m, 2H), 3.90 (s, 2H), 2.46 2.34 (m, 2H), 2.31 (s, 3H), 2.21 (s,
3H). LCMS (ESI+) [M+H]/z Calc'd 507, found 507.
Example 50
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(5-chloro-pyridin-2-yl)-acetamide
##STR00097## was prepared from intermediate 49g and
2-amino-5-chloro pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.47 (d, 1H, J=5.66 Hz), 8.19 (d, 1H, J=2.64
Hz), 7.99 (s, 1H), 7.75 (s, 1H), 7.70 7.67 (m, 1H), 7.45 (d, 1H,
J=8.29 Hz), 7.32 (d, 1H, J=2.45 Hz), 7.15 7.10 (m, 1H), 6.77 6.73
(m, 1H), 4.67 4.56 (m, 2H), 4.22 4.12 (m, 2H), 3.92 (s, 2H), 2.46
2.34(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 514, found 514.
Example 51
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-pyridin-2-yl-acetamide
##STR00098## was prepared from intermediate 49g and 2-aminopyridine
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.47
(d, 1H, J=5.47 Hz), 8.20 (d, 1H, J=4.34 Hz), 7.99 (d, 1H, J=4.34
Hz), 7.75 (s, 1H), 7.72 7.63 (m, 1H), 7.46 (d, 1H, J=8.48 Hz), 7.33
(d, 1H, J=2.26 Hz), 7.18 7.11 (m, 1H), 7.06 6.99 (m, 1H), 6.77 (d,
1H, J=5.46 Hz), 4.65 4.57 (m, 2H), 4.22 4.12(m, 2H), 3.93 (s, 2H),
2.46 2.34(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 479, found 479. Anal.
(C.sub.24H.sub.19N.sub.4O.sub.3SCl.0.5EtOAc.1.2 MeOH)C, H, N.
Example 52
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(5-methyl-isoxazol-3-yl)-acetamide
##STR00099## was prepared from intermediate 49g and
3-amino-5-methyl isoxazole following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.46 (d, 1H, J=4.90 Hz), 7.74 (s, 1H),
7.43 (d, 1H, J=8.29 Hz), 7.31 (d, 1H, J=2.45 Hz), 7.15 (d, 1H,
J=6.79 Hz), 6.75 (d, 1H, J=5.27 Hz), 6.49 (s, 1H), 4.67 4.56 (m,
2H), 4.22 4.12 (m, 2H), 3.86 (s, 2H), 2.46 2.34 (m, 2H), 2.29 (s,
3H). LCMS (ESI+) [M+H]/z Calc'd 483, found 483. Anal.
(C.sub.23H.sub.19N.sub.4O.sub.4SCl.0.6 CH.sub.3COOH.1.5 H.sub.2O)C,
H, N.
Example 53
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(3-methyl-isoxazol-5-yl)-acetamide
##STR00100## was prepared from intermediate 49g and
5-amino-3-methyl isoxazole following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.46 (d, 1H, J=5.28 Hz), 7.74 (s, 1H),
7.43 (d, 1H, J=8.29 Hz), 7.31 (d, 1H, J=2.45 Hz), 7.15 7.10 (m,
1H), 6.75 (d, 1H, J=5.36 Hz), 6.49 (s, 1H), 4.67 4.56 (m, 2H), 4.22
4.12 (m, 2H), 3.87 (s, 2H), 2.46 2.34 (m, 2H), 2.13 (s, 3H). LCMS
(ESI+) [M+H]/z Calc'd 483, found 483.
Example 54
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(4-methyl-oxazol-2-yl)-acetamide
##STR00101## was prepared from intermediate 49g and
2-amino-4-methyl-oxazole following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) 38.47 (d, 1H, J=5.47 Hz), 7.77 (s, 1H), 7.40 (d, 1H,
J=9.23 Hz), 7.33 (d, 1H, J=2.45 Hz), 7.29 (s, 1H), 7.18 7.09 (m,
1H), 6.77 (d, 1H, J=5.09 Hz), 4.67 4.56 (m, 2H), 4.22 4.12 (m, 2H),
3.83 (s, 2H), 2.46 2.34 (m, 2H), 1.97 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 483, found 483.
Example 55
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(5-methyl-1H-pyrazol-3-yl)-acetamide
##STR00102## was prepared from intermediate 49g and
3-amino-5-methyl pyrazole following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.48 (d, 1H, J=5.27 Hz), 7.77 (s, 1H), 7.47 (d,
1H, J=8.67 Hz), 7.32 (s, 1H), 7.14 (d, 1H, J=9.42 Hz), 6.77 (d, 1H,
J=5.27 Hz), 6.22 (s, 1H), 4.67 4.56 (m, 2H), 4.22 4.12 m, 2H), 3.85
(s, 2H), 2.46 2.34 (m, 2H), 2.18 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 482, found 482. Anal.
(C.sub.23H.sub.20N.sub.5O.sub.3SCl.1.7H.sub.2O)C, H, N.
Example 56
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(1,5-dimethyl-1H-pyrazol-3-yl)-acetamide
##STR00103## was prepared from intermediate 49g and
3-amino-1,5-dimethylpyrazole following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.48 (d, 1H, J=5.47 Hz), 7.76 (s, 1H),
7.48 (d, 1H, J=8.29 Hz), 7.34 (d, 1H, J=2.45 Hz), 7.18 7.12 (m,
1H), 6.66 (d, 1H, J=5.46 Hz), 5.97 (s, 1H), 4.67 4.56 (m, 2H), 4.22
4.12 (m, 2H), 3.90 (s, 2H), 3.59 (s, 3H), 2.46 2.34 (m, 2H), 2.10
(s, 3H). LCMS (ESI+) [M+H]/z Calc'd 496, found 496. Anal.
(C.sub.24H.sub.22N.sub.5O.sub.3SCl.0.1CH.sub.2Cl.sub.2.1.0EtOAc) C,
H, N.
Example 57
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-(3-morpholin-4-yl-propyl)-acetamide
##STR00104## was prepared from intermediate 49g and
3-morpholin-4-yl-propylamine following Method A. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.48 (d, 1H, J=5.47 Hz), 7.76 (s, 1H),
7.43 (d, 1H, J=8.48 Hz), 7.31 (d, 1H, J=2.48 Hz), 7.16 7.10 (m,
1H), 6.76 (d, 1H, J=5.46 Hz), 4.67 4.56 (m, 2H), 4.22 4.12 (m, 2H),
3.66 3.59 (m, 6H), 3.24 3.18 (m, 2H), 2.46 2.34(m, 8H), 1.72 1.63
(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 529, found 529. Anal.
(C.sub.26H.sub.29N.sub.4O.sub.4SCl.0.8CH.sub.3COOH) C, H, N.
Example 58
2-{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-
-phenyl}-N-(5-methyl-furan-2-ylmethyl)-acetamide
##STR00105##
Intermediate 58a:
{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]--
phenyl}-acetic acid
##STR00106## was prepared from hydrolysis of its corresponding
methyl ester following the procedure described for the preparation
of intermediate 49g. The corresponding methyl ester was prepared
from coupling of
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine and
49e following Method C. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
8.42 (d, 1H, J=5.66 Hz), 7.72 (s, 1H), 7.42 (d, 1H, J=8.48 Hz),
7.29 (d, 1H, J=2.45 Hz), 7.23 (s, 1H), 7.14 7.08 (m, 1H), 7.10 (s,
1H), 6.70 (d, 1H, J=5.46 Hz), 3.93 (s, 3H), 3.74 (s, 2H). LCMS
(ESI+) [M+H]/z Calc'd 400, found 400.
The compound of Example 58 was prepared from intermediate 58a and
C-(5-methyl-furan-2-yl)-methylamine following Method A. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.37 (d, 1H, J=5.66 Hz), 7.66 (s,
1H), 7.36 (d, 1H, J=8.29 Hz), 7.25 (d, 1H, J=2.26 Hz), 7.19 (s,
1H), 7.10 7.03 (m, 1H), 6.98 (d, 1H, J=1.32 Hz), 6.65(d, 1H, J=5.46
Hz), 6.00(d, 1H, J=3.02 Hz), 5.80 5.78 (m, 1H), 4.22 (s, 2H), 3.90
(s, 3H), 3.64 (s, 2H), 2.12(s, 3H). LCMS (ESI+) [M+H]/z Calc'd 493,
found 493. Anal.
(C.sub.25H.sub.21N.sub.4O.sub.3SCl.0.2CH.sub.2Cl.sub.2), C, H,
N.
Example 59
2-{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-
-phenyl}-N-(3-fluoro-benzyl)-acetamide
##STR00107## was prepared from intermediate 58a and
3-fluoro-benzylamine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.40 (d, 1H, J=5.46 Hz), 7.70 (s, 1H), 7.41 (d,
1H, J=8.47 Hz), 7.28 (d, 1H, J=2.45 Hz), 7.23 7.19 (m, 2H), 7.13
7.07 (m, 1H), 6.99 (d, 1H, J=1.14 Hz), 6.69(d, 1H, J=5.65 Hz), 7.04
6.85 (m, 2H), 6.69(d, 1H, J=5.65 Hz), 4.32 (s, 2H), 3.92 (s, 3H),
3.70 (s, 2H). LCMS (ESI+) [M+H]/z Calc'd 507, found 507. Anal.
(C.sub.26H.sub.20N.sub.4O.sub.2SCl F.0.1CH.sub.2Cl.sub.2), C, H,
N.
Example 60
2-{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-
-phenyl}-N-pyridin-2-yl-acetamide
##STR00108## was prepared from intermediate 58a and 2-aminopyridine
following Method A. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.41
(d, 1H, J=5.65 Hz), 8.22 8.18 (m, 1H), 7.98 (d, 1H, J=8.48 Hz),
7.70 7.63 (m, 2H), 7.46 (d, 1H, J=8.48 Hz), 7.31 (d, 1H, J=2.45
Hz), 7.22 (s, 1H), 7.16 7.10 (m, 1H), 7.05 6.98 (m, 2H), 6.71 (d,
1H, J=5.46 Hz), 3.92 (s, 5H). LCMS (ESI+) [M+H]/z Calc'd 476, found
476. Anal.
(C.sub.24H.sub.18N.sub.5O.sub.2SCl.0.5CH.sub.2Cl.sub.2.0.6EtOAc),
C, H, N.
Example 61
2-{2-Chloro-4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyrid-
in-7-yloxy]-phenyl}-N-pyridin-2-yl-acetamide
##STR00109##
Intermediate 61a:
2-(2-Chloro-4-hydroxy-phenyl)-N-pyridin-2-yl-acetamide
##STR00110## was prepared from (1) coupling of intermediate 49c
with 2-aminopyridine following Method A, and (2) converting the
resulting methyl ether to the corresponding phenol following Method
D. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.16 (d, 1H, J=5.09
Hz), 7.97 (d, 1H, J=8.47 Hz), 7.69 7.60 (m, 1H), 7.10 (d, 1H,
J=8.48 Hz), 7.04 6.97 (m, 1H), 6.76 (d, 1H, J=2.25 Hz), 6.66 6.60
(m, 1H), 3.72 (s, 2H). LCMS (ESI+) [M+H]/z Calc'd 263, found
263.
The compound of Example 61 was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) with intermediate 61a following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.46 (d, 1H, J=5.46 Hz), 8.20 (d, 1H,
J=5.46 Hz), 7.98 (d, 1H, J=8.47 Hz), 7.85 (d, 1H, J=17.33 Hz), 7.70
7.60 (m, 1H), 7.40 (d, 1H, J=8.48 Hz), 7.31 (d, 1H, J=2.45 Hz),
7.16 7.10 (m, 1H), 7.04 6.98 (m, 1H), 6.76 (d, 1H, J=5.46 Hz), 4.41
(bs, 1H), 4.03 3.96 (m, 4H), 3.75 3.57(m, 3H), 2.13 1.94(m, 2H).
LCMS (ESI+) [M+Na]/z Calc'd 510, found 510. Anal.
(C.sub.25H.sub.21N.sub.4O.sub.4SCl.0.5CH.sub.2Cl.sub.2), C, H,
N.
Example 62
2-{2-Chloro-4-[2-((R)-3-methoxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyrid-
in-7-yloxy]-phenyl}-N-pyridin-2-yl-acetamide
##STR00111##
Intermediate 62a
(7-Chloro-thieno[3,2-b]pyridin-2-yl)-(3-methoxy-pyrrolidin-1-yl)-methanon-
e
##STR00112## was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid and
3R-methoxy-pyrrolidine following Method A. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 8.68 (d, 1H, J=5.5 Hz), 7.85 (d, 1H, J=14.3
Hz), 7.40 (d, 1H, J=5.5 Hz), 4.18 4.07 (m, 1H), 4.03 3.73 (m, 4H),
3.20 (d, 3H, J=14.5 Hz), 2.36 2.03 (m, 2H). LCMS ESI (M+H.sup.+):
297.05.
The compound of Example 62 was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-methoxy-pyrrolidin-1-yl)-methanon-
e (62a) with intermediate 61a following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.46 (d, 1H, J=5.46 Hz), 8.20 (d, 1H,
J=6.03 Hz), 7.98 (d, 1H, J=8.10 Hz), 7.84 (d, 1H, J=6.31 Hz), 7.71
7.62 (m, 1H), 7.46 (d, 1H, J=8.48 Hz), 7.31 (d, 1H, J=2.44 Hz),
7.16 7.10 (m, 1H), 7.04 6.98 (m, 1H), 6.66 (d, 1H, J=5.46 Hz), 4.08
3.96 (m, 3H), 3.92 (s, 3H), 3.79 (s, 2H), 3.71 3.57(m, 2H), 2.43
2.33(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 524, found 524. Anal.
(C.sub.26H.sub.23N.sub.4O.sub.4SCl.0.5CH.sub.2Cl.sub.2.1.5EtOAc),
C, H, N.
Example 63
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-2-chloro-phen-
yl}-N-methyl-acetamide
##STR00113##
Intermediate 2-(2-Chloro-4-hydroxy-phenyl)-N-methyl-acetamide
(63a)
##STR00114## was prepared from (1) coupling of intermediate 49c
with methylamine following Method A, and (2) converting methyl
ether to the corresponding phenol following Method D. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 7.02 (d, 1H, J=8.29 Hz), 6.71 (d, 1H,
J=2.45 Hz), 6.61 6.56 (m, 1H), 3.43 (s, 2H), 2.61 (s, 3H). LCMS
(ESI+) [M+H]/z Calc'd 200, found 200.
The compound of Example 63 was prepared from coupling of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
and intermediate 63a following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.47 (d, 1H, J=5.46 Hz), 7.76 (s, 1H), 7.42 (d,
1H, J=8.48 Hz), 7.30 (d, 1H, J=2.45 Hz), 7.15 7.09 (m, 1H), 6.76
(d, 1H, J=5.46 Hz), 4.674.57 (m, 2H), 4.23 4.13(m, 2H), 3.65 (s,
2H), 2.68 (s, 3H), 2.45 2.33 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd
416, found 416. Anal.
(C.sub.20H.sub.18N.sub.3O.sub.3SCl.0.3CH.sub.2Cl.sub.2) C, H,
N.
Example 64
2-{2-Chloro-4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyrid-
in-7-yloxy]-phenyl}-N-methyl-acetamide
##STR00115## was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) with intermediate 63a following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.45 (d, 1H, J=5.47 Hz), 7.84 (d, 1H,
J=17.33 Hz), 7.40 (d, 1H, J=8.48 Hz), 7.29 (d, 1H, J=2.26 Hz), 7.13
7.08 (m, 1H), 6.74 (d, 1H, J=5.46 Hz), 4.42 (bs, 1H), 4.03 3.90 (m,
2H), 3.74 3.58(m, 5H), 2.66 (s, 3H), 2.10 1.97(m, 2H). LCMS (ESI+)
[M+H]/z Calc'd 446, found 446. Anal.
(C.sub.21H.sub.20N.sub.3O.sub.4SCl.0.7CH.sub.2Cl.sub.2), C, H,
N.
Example 65
2-{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-
-phenyl}-N-methyl-acetamide
##STR00116## was prepared from coupling of
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine and
intermediate 63a following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.38 (d, 1H, J=5.65 Hz), 7.67 (s, 1H), 7.38 (d,
1H, J=8.48 Hz), 7.27 (d, 1H, J=2.44 Hz), 7.20 (s, 1H), 7.12 7.06
(m, 1H), 6.99 (s, 1H), 6.67 (d, 1H, J=5.47 Hz), 3.91 (s, 3H),
3.61(s, 2H), 2.66 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 413, found
413. Anal.
(C.sub.20H.sub.17N.sub.4O.sub.2SCl.0.25CH.sub.2Cl.sub.2), C, H,
N.
Example 66
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-methyl-acrylamide
##STR00117##
Intermediate 66a: 2-(4-Methoxy-phenyl)-acrylic acid ethyl ester
##STR00118## A mixture of ethyl-4-methoxyphenethyl acetate (3.0 g,
15.44 mmol), 95% parafomaldehyde (732 mg, 23.16 mmol),
K.sub.2CO.sub.3 (3.30 g, 23.88 mmol), and Bu.sub.4NI (171 mg, 0.463
mmol) in toluene was heated at 80.degree. C. for two hours, cooled
to room temperature, poured into water, and extracted with EtOAc
for three times. The combined organic phase was dried over
Na.sub.2SO.sub.4, concentrated in vacuo, and purified by flash
column chromatography eluting with 10% EtOAc in hexanes to provide
2.14 g of intermediate 66a as a colorless oil. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 7.36 (d, 2H, J=8.67 Hz), 6.87 (d, 2H,
J=8.67 Hz), 6.64 (s, 1H), 5.81 (s, 1H), 4.28 (q, 2H, J=7.16 Hz),
3.81 (s, 3H), 1.32 (t, 2H, J=7.16 Hz). LCMS (ESI+) [M+H]/z Calc'd
207, found 207.
Intermediate 66b: 2-(4-Methoxy-phenyl)-N-methyl-acrylamide
##STR00119## To a solution of 2-(4-methoxy-phenyl)-acrylic acid
ethyl ester (66a) (1.0 g, 4.85 mmol) in 15 mL of THF was added
dropwise 0.33 N KOH (33.5 mL) at 0.degree. C. The mixture was
stirred at ambient temperature for three hours, concentrated in
vacuo, re-suspended in water, and extracted with EtOAc three times.
The combined organic layer was dried over Na.sub.2SO.sub.4, and
concentrated to give 0.86 g acid as a white solid. It was dissolved
in 8 mL of DMF, and to this solution was added 2.0 M methylamine
(9.7 mL, 19.40 mmol) and Et.sub.3N (2.70 mL, 19.40 mmol), follow by
HATU (2.75 g, 7.23 mmol). The resulting mixture was stirred at room
temperature for 30 minutes. Sat. NaHCO.sub.3 was added, the mixture
was extracted with EtOAc. The organic phase was dried over
Na.sub.2SO.sub.4, concentrated, and purified by flash column
chromatography eluting with EtOAc:CH.sub.2Cl.sub.2:MeOH (1:1:0.01)
to provide 0.68 g off-white solid as the intermediate 66b. .sup.1H
NMR (300 MHz, CD.sub.3OD) .delta. 7.23 (d, 2H, J=8.86 Hz), 6.81 (d,
2H, J=8.85 Hz), 5.52 (s, 2H), 3.81 (s, 3H), 2.72 (d, 3H, J=3.77
Hz). LCMS (ESI+) [M+H]/z Calc'd 192, found 192.
Intermediate 66c: 2-(4-Hydroxy-phenyl)-N-methyl-acrylamide
##STR00120## To a solution of
2-(4-methoxy-phenyl)-N-methyl-acrylamide (66b) (0.68 g, 3.56 mmol)
in 40 mL of CH.sub.2Cl.sub.2 was added 1.0 M BBr.sub.3 (7.1 mL,
7.10 mmol) at 0.degree. C. The mixture was stirred at 0.degree. C.
to room temperature for two hours. The reaction was quenched with
MeOH, neutralized with concentrated aqueous NH.sub.4OH to pH
.about.7. The resulting mixture was stirred at room temperature for
one hour. Water was added, and the mixture was extracted with
CH.sub.2Cl.sub.2 three times. The combined organic layer was dried
over Na.sub.2SO.sub.4, concentrated, and purified by flash column
chromatography eluting with EtOAc:CH.sub.2Cl.sub.2:MeOH (1:1:0.02)
to provide 0.38 g orange solid as the intermediate 66c. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 7.17 (d, 2H, J=8.67 Hz), 6.67 (d, 2H,
J=8.85 Hz), 5.48 (d, 2H, J=1.89 Hz), 2.72 (s, 3H). LCMS (ESI+)
[M+H]/z Calc'd 178, found 178.
The compound of Example 66 was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) and intermediate 66c following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.46 Hz), 7.83 (d, 1H,
J=16.96 Hz), 7.46 (d, 2H, J=8.66 Hz), 7.16 (d, 2H, J=8.86 Hz), 6.67
(d, 1H, J=5.46 Hz), 5.70 (d, 2H, J=8.86 Hz), 4.42 (bs, 1H), 3.98
3.88 (m, 2H), 3.71 3.57(m, 3H), 2.75 (s, 3H), 2.13 1.93 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 424, found 424. Anal.
(C.sub.22H.sub.21N.sub.3O.sub.4S.0.6CH.sub.2Cl.sub.2) C, H, N.
Example 67
1-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-cyclopropanecarboxylic acid methylamide
##STR00121##
Intermediate 67a: 1-(4-Methoxy-phenyl)-cyclopropanecarboxylic acid
methylamide
##STR00122## 1-(4-Methoxyphenyl)-cyclopropane carboxyl acid (1.0 g,
5.20 mmol) was dissolved in 6 mL of DMF, to this solution was added
2.0 M methylamine (10.4 mL, 20.80 mmol) and Et.sub.3N (3.0 mL,
20.80 mmol), followed by HATU (3.00 g, 7.89 mmol). The resulting
mixture was stirred at room temperature for 30 min. Saturated
aqueous NaHCO.sub.3 solution was added, and the mixture was
extracted with EtOAc for three times. The combined organic phase
was dried over Na.sub.2SO.sub.4, concentrated, and purified by
flash column chromatography eluting with
EtOAc:CH.sub.2Cl.sub.2:MeOH (1:1:0.02) to provide 0.78 g off-white
solid as the intermediate 67a. .sup.1H NMR (300 MHz, CDCl.sub.3)
.delta. 7.32 (d, 2H, J=8.85 Hz), 6.88 (d, 2H, J=8.66 Hz), 5.38 (bs,
1H), 3.82 (s, 3H), 2.70 (d, 3H, J=4.71 Hz), 1.59 1.54 (m, 2H), 1.00
0.97 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 206, found 206.
Intermediate 67b: 1-(4-Hydroxy-phenyl)-cyclopropanecarboxylic acid
methylamide
##STR00123## was prepared from intermediate 67a following Method D.
.sup.1H NMR (300 MHz, DMSO-d.sub.6) .delta. 9.41 (s, 1H), 7.12 (d,
2H, J=8.47 Hz), 6.71 (d, 2H, J=8.47 Hz), 6.47 (s, 1H), 2.49 (s,
3H), 1.28 1.22 (m, 2H), 0.87 0.80 (m, 2H). LCMS (ESI+) [M+H]/z
Calc'd 192, found 192.
The compound of Example 67 was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) and intermediate 67b following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.28 Hz), 7.82 (d, 1H,
J=17.52 Hz), 7.43 (d, 2H, J=8.47 Hz), 7.15 (d, 2H, J=8.47 Hz), 6.76
(d, 1H, J=5.46 Hz), 4.42 (bs, 1H), 4.01 3.88 (m, 2H), 3.76 3.57(m,
3H), 2.59 (s, 3H), 2.13 1.93 (m, 2H), 1.46 1.39 (m, 2H), 1.02 0.96
(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 438, found 438. Anal.
(C.sub.23H.sub.23N.sub.3O.sub.4S.0.3CH.sub.2Cl.sub.2) C, H, N.
Example 68
2-{4-[2-((S)-3-Methoxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-methyl-acetamide
##STR00124##
Intermediate 68a: 2-(4-Methoxy-phenyl)-N-methyl-acetamide
##STR00125## was prepared from 4-methoxyphenylacetic acid chloride
and methylamine following Method B. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.14 (d, 2H, J=8.67 Hz), 6.86 (d, 2H, J=8.67
Hz), 3.78 (s, 3H), 3.49 (s, 2H), 2.72 (d, 3H, J=4.71 Hz). LCMS
(ESI+) [M+H]/z Calc'd 180, found 180.
Intermediate 68b: 2-(4-Hydroxy-phenyl)-N-methyl-acetamide
##STR00126## was prepared from intermediate 68a following Method D.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 6.97 (d, 2H, J=8.48 Hz),
6.61 (d, 2H, J=8.67 Hz), 3.27 (s, 2H), 2.59 (d, 3H, J=4.52 Hz).
LCMS (ESI+) [M+H]/z Calc'd 166, found 166.
Intermediate 68c:
(7-chloro-thien[3,2-b]pyridin-2-yl)-(3-(S)-methoxy-pyrrolidin-1-yl)-metha-
none
##STR00127## was prepared from
7-chloro-thieno[3,2-b]pyridine-2-carboxylic acid and
3S-methoxy-pyrrolidine following Method A. .sup.1H NMR (300 MHz,
CDCl.sub.3, .delta. 8.68 (d, 1H, J=5.5 Hz), 7.85 (d, 1H, J=14.3 Hz,
7.40 (d, 1H, J=5.5 Hz), 4.18 4.07 (m, 1H), 4.03 3.73 (m, 4H), 3.2
(d, 3H, J=14.5 Hz), 2.36 2.03 (m, 2H). LCMS ESI (M+H+): 297.05
The compound of Example 68 was prepared from
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3(S)-methoxy-pyrrolidin-1-yl)-metha-
none (68c) and intermediate 68b following Method C. .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.39 (d, 1H, J=5.46 Hz), 7.80 (d, 1H,
J=4.53 Hz), 7.34 (d, 2H, J=8.48 Hz), 7.10 (d, 2H, J=8.67 Hz), 6.60
(d, 1H, J=5.47 Hz), 4.05 3.80 (m, 3H), 3.70 3.58 (m, 2H), 3.46 (s,
2H), 3.29 (s, 3H), 2.65 (s, 3H), 2.11 1.95 (m, 2H). LCMS (ESI+)
[M+H]/z Calc'd 426, found 426. Anal.
(C.sub.22H.sub.23N.sub.3O.sub.4S.0.2CH.sub.2Cl.sub.2) C, H, N.
Example 69
2-{4-[2-((R)-3-Hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-methyl-acetamide
##STR00128## was prepared from
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) and intermediate 68b following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.39 (d, 1H, J=5.65 Hz), 7.81 (d, 1H,
J=17.14 Hz), 7.43 (d, 2H, J=8.48 Hz), 7.10 (d, 2H, J=8.67 Hz), 6.61
(d, 1H, J=5.46 Hz), 4.42 (bs, 1H), 4.01 3.88 (m, 2H), 3.76 3.57(m,
3H), 3.45 (s, 2H), 2.64 (s, 3H), 2.09 1.98 (m, 2H). LCMS (ESI+)
[M+H]/z Calc'd 412, found 412. Anal.
(C.sub.21H.sub.21N.sub.3O.sub.4S.0.4CH.sub.2Cl.sub.2) C, H, N.
Example 70
2-{2-Chloro-4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyrid-
in-7-yloxy]-phenyl}-2-methoxy-N-methyl-acetamide
##STR00129##
Intermediate 70a:
2-(2-Chloro-4-methoxy-phenyl)-N-methyl-acetamide
##STR00130## To a suspension of (2-chloro-4-methoxy-phenyl)-acetic
acid (49c) (500 mg, 2.50 mmol) in CH.sub.2Cl.sub.2 (10 mL) was
added 2.0 M oxalyl chloride in CH.sub.2Cl.sub.2 (3.75 mL, 7.50
mmol), follow by 4 drops of DMF. The mixture was stirred at ambient
temperature for one hour, concentrated, and further dried under
high vacuum. The residue was re-dissolved in CH.sub.2Cl.sub.2 (10
mL), to this solution was added 2.0 M methylamine in THF (3.5 mL,
7.50 mmol). After stirring at ambient temperature overnight, the
reaction was quenched with water, extracted with CH.sub.2Cl.sub.2
for three times. The combined organic phase was dried over
Na.sub.2SO.sub.4, and concentrated to give 0.46 g white solid as
the intermediate 70a. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.14 (d, 1H, J=8.66 Hz), 6.87 (d, 1H, J=2.64 Hz), 6.78 6.72 (m,
1H), 3.68 (s, 3H), 3.48 (s, 2H), 2.62 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 214, found 214.
Intermediate 70b:
2-Bromo-2-(2-chloro-4-methoxy-phenyl)-N-methyl-acetamide
##STR00131## A mixture of
2-(2-chloro-4-methoxy-phenyl)-N-methyl-acetamide (70a) (0.22 g,
1.03 mmol), NBS (183 mg, 1.03 mmol), and 70% benzyl peroxide (36
mg, 0.103 mmol) in 6 mL of CCl.sub.4 was refluxed at 80.degree. C.
for three hours, cooled to room temperature, filtered, and
concentrated in vacuo. The residue was purified by flash column
chromatography eluting with 1:1 EtOAc and hexanes to provide 185 mg
white solid as the intermediate 70b. .sup.1H NMR (300 MHz,
CDCl.sub.3) .delta. 7.42 (d, 1H, J=8.85 Hz), 6.90 (d, 1H, J=2.63
Hz), 6.83 6.76 (m, 1H), 5.83 (s, 1H), 3.78 (s, 3H), 2.90 (d, 3H,
J=4.90 Hz). LCMS (ESI+) [M+H]/z Calc'd 294, found 294.
Intermediate 70c:
2-(2-Chloro-4-hydroxy-phenyl)-2-methoxy-N-methyl-acetamide
##STR00132## To a solution of
2-bromo-2-(2-chloro-4-methoxy-phenyl)-N-methyl-acetamide (70b) (185
mg, 0.632 mmol) in 5 mL of CH.sub.2Cl.sub.2 was added 1.0 M
BBr.sub.3 (1.90 mL, 1.90 mmol) in CH.sub.2Cl.sub.2 at 0.degree. C.
The mixture was stirred at 0.degree. C. to room temperature for two
hours. The reaction was quenched with MeOH, neutralized with
concentrated aqueous NH.sub.4OH. The resulting mixture was stirred
at room temperature for one hour. Water was added, the mixture was
extracted with CH.sub.2Cl.sub.2 for three times. The combined
organic layer was dried over Na.sub.2SO.sub.4, concentrated, and
purified by flash column chromatography eluting with 3:1 EtOAc and
hexanes to provide 80 mg white solid as the intermediate 70c.
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.06 (s, 1H), 7.06 (d,
1H, J=8.48 Hz), 6.73 (d, 1H, J=2.45 Hz), 6.65 6.59 (m, 1H), 4.91
(s, 1H), 3.22 (s, 3H), 2.69 (s, 3H, J=4.71 Hz). LCMS (ESI+) [M+H]/z
Calc'd 230, found 230.
The compound of Example 70 was prepared from the coupling of
(7-chloro-thieno[3,2-b]
pyridin-2-yl)-((R)-3-hydroxy-pyrrolidin-1-yl)-methanone (2b) and
intermediate 70c following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.46 (d, 1H, J=5.46 Hz), 7.85 (d, 1H, J=17.33
Hz), 7.45 (d, 1H, J=8.47 Hz), 7.31 (d, 1H, J=2.26 Hz), 7.18 7.13
(m, 1H), 6.73 (d, 1H, J=5.46 Hz), 5.07 (s, 1H), 4.42 (bs, 1H), 4.03
3.90 (m, 2H), 3.74 3.58 (m, 3H), 3.31 (s, 3H), 2.72 (s, 3H), 2.06
1.97 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 476, found 476. Anal.
(C.sub.22H.sub.22N.sub.3O.sub.5SCl. 0.2CH.sub.2Cl.sub.2.0.2EtOAc),
C, H, N.
Example 71
2-{2-Chloro-4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyrid-
in-7-yloxy]-phenyl}-2-hydroxy-N-methyl-acetamide
##STR00133## was prepared from the title compound of Example 70
following Method D. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.46
(d, 1H, J=5.46 Hz), 7.85 (d, 1H, J=17.52 Hz), 7.46 (d, 1H, J=8.48
Hz), 7.28 (d, 1H, J=2.45 Hz), 7.16 7.08 (m, 1H), 6.71 (d, 1H,
J=5.46 Hz), 5.41 (s, 1H), 4.42 (bs, 1H), 4.03 3.90 (m, 3H), 3.74
3.58(m, 2H), 2.73 (s, 3H), 2.12 1.97(m, 2H). LCMS (ESI+) [M+H]/z
Calc'd 462, found 462 Anal.
(C.sub.21H.sub.20N.sub.3O.sub.5SCl.0.7CH.sub.2Cl.sub.2) C, H,
N.
Example 72
2-{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-
-phenyl}-2-methoxy-N-methyl-acetamide
##STR00134## was prepared from the coupling of
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine and
intermediate 70c following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.46 Hz), 7.71 (s, 1H), 7.48 (d,
1H, J=8.47 Hz), 7.34 (d, 1H, J=2.26 Hz), 7.24 (s, 1H), 7.21 7.15
(m, 1H), 7.02 (s, 1H), 6.69 (d, 1H, J=5.46 Hz), 5.09 (s, 1H), 3.94
(s, 3H), 3.34 (s, 3H), 2.75 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd
443, found 443. Anal.
(C.sub.21H.sub.19N.sub.4O.sub.3SCl.0.7CH.sub.2Cl.sub.2.0.3EtOAc) C,
H, N.
Example 73
2-{2-Chloro-4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy]-
-phenyl}-2-hydroxy-N-methyl-acetamide
##STR00135## was prepared from the title compound of Example 72
following Method D. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.41
(d, 1H, J=5.46 Hz), 7.70 (s, 1H), 7.46 (d, 1H, J=8.47 Hz), 7.28 (d,
1H, J=2.45 Hz), 7.21 (s, 1H), 7.16 7.10 (m, 1H), 7.00 (s, 1H), 6.67
(d, 1H, J=5.65 Hz), 5.39 (s, 1H), 3.92 (s, 3H), 2.73 (s, 3H). LCMS
(ESI+) [M+H]/z Calc'd 429, found 429. Anal.
(C.sub.20H.sub.17N.sub.4O.sub.3SCl.0.5EtOAc.0.5MeOH)C, H, N.
Example 74
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-2-hyd-
roxy-N-methyl-acetamide
##STR00136##
Intermediate 74a: 2-(4-Methoxy-phenyl)-N-methyl-2-oxo-acetamide
##STR00137## To a solution of ethyl-4-methoxylbenzoformate (2.00 g,
7.99 mmol) in 17 mL of THF was added dropwise 0.33 N KOH (55 mL) at
0.degree. C. The mixture was stirred at room temperature for three
hours, and concentrated in vacuo. Saturated aqueous NaHCO.sub.3
solution was added, and the mixture was extracted with EtOAc. The
combined organic layer was dried over Na.sub.2SO.sub.4, and
concentrated to give 1.969 acid as white solid. It was dissolved in
10 mL of DMF, and to this solution was added 2.0 M methylamine in
THF (21.78 mL, 43.56 mmol) and Et.sub.3N (6.07 mL, 43.56 mmol),
follow by HATU (6.71 g, 17.65 mmol). The resulting mixture was
stirred at room temperature for one hour. Water was added, the
mixture was extracted with EtOAc. The organic phase was dried over
Na.sub.2SO.sub.4, concentrated, and purified by flash column
chromatography eluting with EtOAc:CH.sub.2Cl.sub.2:MeOH (1:1:0.01)
to provide 1.35 g off-white solid as the intermediate 74a. .sup.1H
NMR (300 MHz, CD.sub.3OD) .delta. 8.03 (d, 2H, J=9.04 Hz), 6.94 (d,
2H, J=9.04 Hz), 3.80 (s, 3H), 2.78 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 194, found 194.
Intermediate 74b: 2-(4-Hydroxy-phenyl)-N-methyl-2-oxo-acetamide
##STR00138## To a solution of
2-(4-methoxy-phenyl)-N-methyl-2-oxo-acetamide (74a) (1.35 g, 6.99
mmol) in 25 mL of CH.sub.2Cl.sub.2 was added 1.0 M BBr.sub.3 in
CH.sub.2Cl.sub.2 (14 mL, 28.0 mmol) at 0.degree. C. The mixture was
stirred at 0.degree. C. to room temperature for two hours. The
reaction was quenched with MeOH, neutralized with concentrated
aqueous NH.sub.4OH to pH .about.7. The resulting mixture was
stirred at room temperature for one hour. Water was added, and the
mixture was extracted with CH.sub.2Cl.sub.2. The organic phase was
dried over Na.sub.2SO.sub.4, concentrated, and purified by flash
column chromatography eluting with 1:1 EtOAc and hexanes to provide
0.53 g off-white solid as the intermediate 74b. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 7.03 (d, 2H, J=8.86 Hz), 6.75 (d, 2H,
J=8.86 Hz), 2.76 (s, 3H). LCMS (ESI+) [M+Na]/z Calc'd 202, found
202.
Intermediate 74c:
2-Hydroxy-2-(4-hydroxy-phenyl)-N-methyl-acetamide
##STR00139## To solution of
2-(4-hydroxy-phenyl)-N-methyl-2-oxo-acetamide (74b) (121 mg, 0.676
mmol) in 3 mL of MeOH was added NaBH.sub.4 (51 mg, 1.352 mmol). The
mixture was stirred at room temperature for 30 min, and
concentrated. 1.0 N aqueous HCl solution was added, the mixture was
extracted with EtOAc for three times. The organic phase was dried
over Na.sub.2SO.sub.4, and concentrated to give 114 mg of the
intermediate 74c as a white solid. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.18 (d, 2H, J=8.48 Hz), 6.67 (d, 2H, J=8.48
Hz), 4.78 (s, 1H), 2.61 (s, 3H). LCMS (ESI+) [M+Na]/z Calc'd 204,
found 204.
The compound of Example 74 was prepared from coupling of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
and intermediate 74c following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.47 Hz), 7.72 (s, 1H), 7.52 (d,
2H, J=8.48 Hz), 7.14 (d, 2H, J=8.48 Hz), 6.61 (d, 1H, J=5.47 Hz),
4.97 (s, 1H), 4.67 4.57 (m, 2H), 4.23 4.13(m, 2H), 2.69 (s, 3H),
2.45 2.33 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 398, found 398. Anal.
(C.sub.20H.sub.19N.sub.3O.sub.4S.0.6CH.sub.2Cl.sub.2) C, H, N.
Example 75
2-Hydroxy-2-(4-{2-[4-(1-hydroxy-1-methyl-ethyl)-thiazol-2-yl]-thieno[3,2-b-
]pyridin-7-yloxy}-phenyl)-N-methyl-acetamide
##STR00140## was prepared from the coupling of
2-[2-(7-chloro-thieno[3,2-b]pyridin-2-yl)-thiazol-4-yl]-propan-2-ol
and intermediate 74c following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.36 (d, 1H, J=5.46 Hz), 7.82 (s, 1H), 7.61 (d,
2H, J=8.66 Hz), 7.39 (s, 1H), 7.15 (d, 2H, J=8.66 Hz), 6.61 (d, 1H,
J=5.47 Hz), 4.98 (s, 1H), 2.69 (s, 3H), 1.52 (s, 6H). LCMS (ESI+)
[M+H]/z Calc'd 456, found 456. Anal.
(C.sub.22H.sub.21N.sub.3O.sub.4S.sub.2.0.2CH.sub.2Cl.sub.2) C, H,
N.
Example 76
2-Hydroxy-2-{4-[2-((R)-3-hydroxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyri-
din-7-yloxy]-phenyl}-N-methyl-acetamide
##STR00141## was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-hydroxy-pyrrolidin-1-yl)-methanon-
e (2b) and intermediate 74c following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.47 Hz), 7.84 (d, 1H,
J=17.33 Hz), 7.52 (d, 2H, J=8.48 Hz), 7.14 (d, 2H, J=8.48 Hz), 6.61
(d, 1H, J=5.47 Hz), 5.09 (s, 1H), 4.42 (bs, 1H), 4.23 3.90 (m, 2H),
3.89 3.54(m, 3H), 2.69 (s, 3H), 2.20 1.97(m, 2H). LCMS (ESI+)
[M+H]/z Calc'd 428, found 428. Anal.
(C.sub.21H.sub.21N.sub.3O.sub.5S.0.6CH.sub.2Cl.sub.2.1.0H.sub.2O)C,
H, N.
Example 77
2-Hydroxy-2-{4-[2-((R)-3-methoxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyri-
din-7-yloxy]-phenyl}-N-methyl-acetamide
##STR00142## was prepared from coupling of
(7-chloro-thieno[3,2-b]pyridin-2-yl)-(3-methoxy-pyrrolidin-1-yl)-methanon-
e (62a) and intermediate 74c following Method C. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.41 (d, 1H, J=5.47 Hz), 7.82 (s, 1H),
7.52 (d, 2H, J=8.48 Hz), 7.14 (d, 2H, J=8.48 Hz), 6.61 (d, 1H,
J=5.47 Hz), 4.98 (s, 1H), 4.08 3.76 (m, 5H), 3.75 3.52 (m, 2H),
2.69 (s, 3H), 2.33 1.97(m, 2H). LCMS (ESI+) [M+H]/z Calc'd 442,
found 442. Anal. (C.sub.22H.sub.23N.sub.3O.sub.5S.2.0H.sub.20)C, H,
N.
Example 78
2-{4-[2-((R)-3-Methoxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyridin-7-ylox-
y]-phenyl}-N-methyl-2-oxo-acetamide
##STR00143## To a solution of Dess-Martin reagent (48 mg, 0.114
mmol) in 1.5 mL of CH.sub.2Cl.sub.2 cooled to 0.degree. C., was
added dropwise a solution of
2-hydroxy-2-{4-[2-((R)-3-methoxy-pyrrolidine-1-carbonyl)-thieno[3,2-b]pyr-
idin-7-yloxy]-phenyl}-N-methyl-acetamide (Example 77) (42 mg, 0.095
mmol) in 1.5 mL of CH.sub.2Cl.sub.2. The mixture was stirred at
0.degree. C. to room temperature for one hour. Saturated aqueous
NaHCO.sub.3 solution was added, and the mixture was extracted with
CH.sub.2Cl.sub.2 and a small amount of MeOH. The combined organic
phase was dried over Na.sub.2SO.sub.4, concentrated, and purified
by flash column chromatography eluting with
EtOAc:CH.sub.2Cl.sub.2:MeOH (1:1:0.04) to provide 30 mg white solid
as the compound of Example 78. .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 8.47 (d, 1H, J=5.47 Hz), 8.17 (d, 2H, J=8.66 Hz), 7.28 (d,
2H, J=8.66 Hz), 7.78 7.62 (m, 1H), 6.81 (d, 1H, J=5.47 Hz), 4.08
3.76 (m, 5H), 3.75 3.52 (m, 3H), 2.78 (s, 3H), 2.30 1.97 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 440, found 440. Anal.
(C.sub.22H.sub.21N.sub.3O.sub.5S.0.4CH.sub.2Cl.sub.2) C, H, N.
Example 79
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-2-met-
hoxy-N-(3-methyl-isoxazol-5-yl)-acetamide
##STR00144##
Intermediate 79a: acetoxy-(4-methoxy-phenyl)-acetic acid
##STR00145## 4-Methoxymandelic acid (5.00 g, 27.44 mmol) was
dissolved in 100 mL of THF. To this solution was added acetic
anhydride (2.85 mL, 30.18 mmol), followed by Et.sub.3N (10 mL). The
resulting mixture was stirred at room temperature overnight,
concentrated, re-suspended in water, and extracted with EtOAc. The
combined organic phase was dried over Na.sub.2SO.sub.4, and
concentrated to give 5.95 g yellow syrup as the intermediate 79a.
.sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.38 (d, 2H, J=8.67 Hz),
6.90 (d, 2H, J=8.67 Hz), 5.87 (s, 1H), 3.80 (s, 3H), 2.16 (s, 3H).
LCMS (ESI-) [M-H]/z Calc'd 223, found 223.
Intermediate 79b: acetic acid
(4-methoxy-phenyl)-(3-methyl-isoxazol-5-ylcarbamoyl)-methyl
ester
##STR00146## To a suspension of acetoxy-(4-methoxy-phenyl)-acetic
acid (79a) (2.78 g, 12.40 mmol) in CH.sub.2Cl.sub.2 (50 mL), was
added 2.0 M oxalyl chloride in CH.sub.2Cl.sub.2 (9.30 mL, 18.60
mmol), followed by 10 drops of DMF. The mixture was stirred at
ambient temperature for one hour, concentrated and further dried
under high vacuum. It was re-dissolved in CH.sub.2Cl.sub.2 (50 mL);
to this solution was added 5-amino-3-methylisoxazole (1.34 g, 13.66
mmol), followed by Et.sub.3N (2.60 mL, 18.60 mmol). After stirring
at ambient temperature overnight, the mixture was concentrated in
vacuo and purified by flash column chromatography eluting with 1:1
EtOAc and hexanes to provide 2.70 g yellow solid as the
intermediate 79b. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 8.56
(s, 1H), 7.36 (d, 2H, J=8.67 Hz), 6.90 (d, 2H, J=8.67 Hz), 6.24 (s,
1H), 6.17 (s, 1H), 3.80 (s, 3H), 2.25 (s, 3H), 2.22 (s, 3H). LCMS
(ESI--) [M-H]/z Calc'd 303, found 303.
Intermediate 79c:
2-(4-Hydroxy-phenyl)-2-methoxy-N-(3-methyl-isoxazol-5-yl)-acetamide
##STR00147## To a solution of acetic acid
(4-methoxy-phenyl)-(3-methyl-isoxazol-5-ylcarbamoyl)-methyl ester
(79b) (1.07 g, 3.52 mmol) in 40 mL of CH.sub.2Cl.sub.2, was added
1.0 M BBr.sub.3 in CH.sub.2Cl.sub.2 (8.8 mL, 8.80 mmol) at
0.degree. C. The mixture was stirred at 0.degree. C. to room
temperature for two hours. The reaction was quenched with MeOH, and
then neutralized with concentrated aqueous NH.sub.4OH. The
resulting mixture was stirred at room temperature for one hour.
Water was added. The mixture was extracted with CH.sub.2Cl.sub.2.
The combined organic layer was dried over Na.sub.2SO.sub.4,
concentrated, and purified by flash column chromatography eluting
with 1:1 EtOAc and hexanes to provide 0.57 g yellow solid as the
intermediate 79c. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.53
(d, 2H, J=7.53 Hz), 6.83 (d, 2H, J=7.54 Hz), 6.25 (s, 1H), 4.90 (s,
1H), 3.40 (s, 3H), 2.25 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 263,
found 263.
The compound of Example 79 was prepared from coupling of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
and intermediate 79c following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.41 (d, 1H. J=5.46 Hz), 7.70 (s, 1H), 7.55 (d,
2H, J=8.67 Hz), 7.20 (d, 2H, J=8.47 Hz), 6.62 (d, 1H, J=5.46 Hz),
6.14 (s, 1H), 4.87 (s, 1H), 4.63 4.52 (m, 2H), 4.21 4.09 (m, 2H),
3.39 (s, 3H), 2.43 2.29 (m, 2H), 2.14 (s, 3H). LCMS (ESI+) [M+H]/z
Calc'd 479, found 479. Anal.
(C.sub.24H.sub.22N.sub.4O.sub.5S.0.1CH.sub.2Cl.sub.2) C, H, N.
Example 80
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-2-hyd-
roxy-N-(3-methyl-isoxazol-5-yl)-acetamide
##STR00148## was prepared from the title compound of Example 79
following Method D. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.43
(d, 1H, J=5.65 Hz), 7.74 (s, 1H), 7.60(d, 2H, J=8.67 Hz), 7.19 (d,
2H, J=8.48 Hz), 6.45 (d, 1H, J=5.46 Hz), 6.16 (s, 1H), 5.21 (s,
1H), 4.63 4.52 (m, 2H), 4.21 4.09 (m, 2H), 2.43 2.29 (m, 2H), 2.15
(s, 3H). LCMS (ESI+) [M+H]/z Calc'd 465, found 465. Anal.
(C.sub.23H.sub.20N.sub.4O.sub.5S.0.4CH.sub.2Cl.sub.2) C, H, N.
Example 81
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-but-
yl-2-methoxy-acetamide
##STR00149##
Intermediate 81a: Acetic acid
butylcarbamoyl-(4-methoxy-phenyl)-methyl ester
##STR00150## was prepared from intermediate 79a and butylamine
following Method A. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 7.34
(d, 2H, J=8.85 Hz), 6.87 (d, 2H, J=8.67 Hz), 6.01 (s, 1H), 3.79 (s,
3H), 3.31 3.23 (m, 2H), 2.15 (s, 3H), 1.54 1.43 (m, 2H), 1.37 1.22
(m, 2H), 0.94 0.87 (m, 3H). LCMS (ESI+) [2M+Na]/z Calc'd 581, found
581.
Intermediate 81 b:
N-Butyl-2-(4-hydroxy-phenyl)-2-methoxy-acetamide
##STR00151## was prepared from intermediate 81a following a similar
procedure as in the conversion of 79b to 79c. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.06(d, 2H, J=8.67 Hz), 6.62 (d, 2H, J=8.66
Hz), 4.36 (s, 1H), 3.15 (s, 3H), 3.10 3.02 (m, 2H), 1.40 1.28 (m,
2H), 1.23 1.09 (m, 2H), 0.80 0.70 (m, 3H). LCMS (ESI+) [M+H]/z
Calc'd 238, found 238.
The compound of Example 81 was prepared from coupling of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone and
intermediate 81b following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.43 (d, 1H, J=5.46 Hz), 7.74 (s, 1H), 7.53(d,
2H, J=8.66 Hz), 7.16 (d, 2H, J=8.48 Hz), 6.63 (d, 1H, J=5.47 Hz),
4.62 (s, 1H), 4.62 4.54 (m, 2H), 4.19 4.00 (m, 2H), 3.33 (s, 3H),
3.21 3.09 (m, 2H), 2.44 2.30 (m, 2H), 1.49 1.36 (m, 2H), 1.31 1.18
(m, 2H), 0.89 0.77 (m, 3H). LCMS (ESI+) [M+H]/z Calc'd 454, found
454. Anal. (C.sub.24H.sub.27N.sub.3O.sub.4S.0.1CH.sub.2Cl.sub.2) C,
H. N.
Example 82
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-N-but-
yl-2-hydroxy-acetamide
##STR00152## was prepared from the title compound of Example 81
following Method D. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.40
(d, 1H, J=5.46 Hz), 7.67 (s, 1H), 7.49(d, 2H, J=8.47 Hz), 7.18 (d,
2H, J=8.48 Hz), 6.61 (d, 1H, J=5.47 Hz), 4.99 (s, 1H), 4.66 4.56
(m, 2H), 4.22 4.12 (m, 2H), 3.21 3.12 (m, 2H), 2.46 2.36 (m, 2H),
1.49 1.36 (m, 2H), 1.31 1.18 (m, 2H), 0.89 0.77 (m, 3H). LCMS
(ESI+) [M+H]/z Calc'd 440, found 440. Anal.
(C.sub.23H.sub.25N.sub.3O.sub.4S.0.2CH.sub.2Cl.sub.2) C, H, N.
Example 83
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-2-met-
hoxy-N-pyridin-2-yl-acetamide
##STR00153##
Intermediate 83a: Acetic acid
(4-methoxy-phenyl)-(pyridin-2-ylcarbamoyl)-methyl ester
##STR00154## was prepared from intermediate 79a and 2-aminopyridine
following Method A. .sup.1H NMR (300 MHz, CDCl.sub.3) .delta. 9.06
(s, 1H), 8.31 8.23 (m, 2H), 7.81 7.75 (m, 1H), 7.43 (d, 2H, J=8.59
Hz), 6.90 (d, 2H, J=8.59 Hz), 6.14 (s, 1H), 3.79 (s, 3H), 2.25 (s,
3H). LCMS (ESI+) [M+H]/z Calc'd 301, found 301.
Intermediate 83b:
2-(4-Hydroxy-phenyl)-2-methoxy-N-pyridin-2-yl-acetamide
##STR00155## was prepared from intermediate 83a following a similar
procedure as in the conversion of intermediate 79b to intermediate
79c. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.21 (d, 1H, J=5.05
Hz), 8.01 (d, 1H J=8.34 Hz), 7.72 7.66 (m, 1H), 7.20 (d, 2H, J=8.59
Hz), 7.08 7.01 (m, 1H), 6.72 (d, 2H, J=8.34 Hz), 4.66 (s, 1H), 3.31
(s, 3H). LCMS (ESI+) [M+H]/z Calc'd 259, found 259.
The compound of Example 83 was prepared from coupling of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
and intermediate 83b following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.38 (d, 1H, J=5.28 Hz), 8.24 8.15 (m, 1H),
8.00 (d, 1H, J=8.29 Hz), 7.72 7.62 (m, 2H), 7.55 (d, 2H, J=8.29
Hz), 7.18 (d, 2H, J=8.29 Hz), 7.09 6.99 (m, 1H), 6.61 (d, 1H,
J=5.27 Hz), 5.39 (s, 1H), 4.63 4.48 (m, 2H), 4.21 4.08 (m, 2H),
3.40 (s, 3H), 2.45 2.28 (m, 2H). LCMS (ESI+) [M+H]/z Calc'd 475,
found 475. Anal.
(C.sub.25H.sub.22N.sub.4O.sub.4S.0.4EtOAc.0.3CH.sub.2Cl.sub.2) C,
H, N.
Example 84
2-{4-[2-(Azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl}-2-hyd-
roxy-N-pyridin-2-yl-acetamide
##STR00156## was prepared from the title compound of Example 83
following Method D. .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 8.42
(d, 1H, J=5.46 Hz), 8.26 8.20 (m, 1H), 8.06 (d, 1H, J=8.29 Hz),
7.77 7.69 (m, 2H), 7.63 (d, 2H, J=8.67 Hz), 7.19 (d, 2H, J=8.66
Hz), 7.11 7.03 (m, 1H), 6.65 (d, 1H, J=5.65 Hz), 5.20 (s, 1H), 4.65
4.55 (m, 2H), 4.23 4.13 (m, 2H), 2.47 2.33 (m, 2H). LCMS (ESI+)
[M+H]/z Calc'd 461, found 461. Anal.
(C.sub.24H.sub.20N.sub.4O.sub.4S.0.7EtOAc.0.5CH.sub.2Cl.sub.2) C,
H, N.
Example 85
2-Methoxy-2-{4-[2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridin-7-yloxy-
]-phenyl}-N-pyridin-2-yl-acetamide
##STR00157## was prepared from coupling of
7-chloro-2-(1-methyl-1H-imidazol-2-yl)-thieno[3,2-b]pyridine and
intermediate 83b following Method C. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.44 (d, 1H, J=5.31 Hz), 8.00 (d, 2H, J=9.10
Hz), 7.78 7.74 (m, 1H), 7.72 (s, 1H), 7.38 7.32 (m, 1H), 7.28 7.20
(m, 3H), 7.00 (s, 1H), 6.61 (d, 1H, J=5.27 Hz), 6.51 6.45 (m, 2H),
3.93 (s, 3H), 3.83 (s, 3H). LCMS (ESI+) [M+H]/z Calc'd 472, found
472. Anal.
(C.sub.25H.sub.21N.sub.5O.sub.3S.0.5EtOAc.1.0CH.sub.2Cl.sub.2) C,
H, N.
Example 86
butyl-carbamic acid
4-[2-(azetidine-1-carbonyl)-thieno[3,2-b]pyridin-7-yloxy]-phenyl
ester
##STR00158##
Intermediate 86a:
Azetidin-1-yl-[7-(4-methoxy-phenoxy)-thieno[3,2-b]pyridin-2-yl]-methanone
##STR00159## A mixture of
azetidin-1-yl-(7-chloro-thieno[3,2-b]pyridin-2-yl)-methanone (21a)
(200 mg, 0.794 mmol), 4-methoxyphenol (148 mg, 1.191 mmol), and
Cs.sub.2CO.sub.3 (391 mg, 1.191 mmol) in 2 mL of DMSO was heated at
100.degree. C. for overnight. It was cooled to room temperature,
EtOAc and 1.0 N NaOH were added. The organic layer was washed with
water and brine, dried over Na.sub.2SO.sub.4, and concentrated to
give 0.27 g of the intermediate 86a as off-white solid. .sup.1H NMR
(300 MHz, CDCl.sub.3) .delta. 8.46 (d, 1H, J=5.46 Hz), 7.75 (s,
1H), 7.09 (d, 2H, J=9.05 Hz), 6.94 (d, 2H, J=9.04 Hz), 6.53 (d, 1H,
J=5.46 Hz), 4.63 4.53 (m, 2H), 4.31 4.20 (m, 2H), 2.40 2.36 (m,
2H). LCMS (ESI+) [M+H]/z Calc'd 341, found 341.
Intermediate 86b:
azetidin-1-yl-[7-(4-hydroxy-phenoxy)-thieno[3,2-b]pyridin-2-yl]-methanone
##STR00160## To a solution of
azetidin-1-yl-[7-(4-methoxy-phenoxy)-thieno[3,2-b]pyridin-2-yl]-methanone
(86a) (400 mg, 1.176 mmol) in 10 mL of CH.sub.2Cl.sub.2 was added
1.0 M BBr.sub.3 (3.53 mL, 3.53 mmol). The mixture was stirred at
room temperature overnight. The reaction was quenched with MeOH,
and then neutralized with concentrated aqueous NH.sub.4OH to pH
.about.7. The resulting mixture was stirred at room temperature for
one hour. Water was added; the mixture was extracted with
CH.sub.2Cl.sub.2. The combined organic layer was dried over
Na.sub.2SO.sub.4, concentrated, and purified by flash column
chromatography eluting with EtOAc:Hex:MeOH (1:1:0.01) to provide
187 mg off-white solid as the intermediate 86b. .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 8.39 (d, 1H, J=5.56 Hz), 7.70 (s, 1H),
6.97 (d, 2H, J=8.84 Hz), 6.80 (d, 2H, J=8.84 Hz), 6.57 (d, 1H,
J=5.56 Hz), 4.65 4.54 (m, 2H), 4.23 4.11(m, 2H), 2.39 2.33 (m, 2H).
LCMS (ESI+) [M+H]/z Calc'd 327, found 327.
To a suspension of
azetidin-1-yl-[7-(4-hydroxy-phenoxy)-thieno[3,2-b]pyridin-2-yl]-methanone
(86b) (41 mg, 0.126 mmol) in 3 mL of toluene, was added 20%
phosgene in toluene (1.38 mL). The mixture was stirred at room
temperature for 15 minutes, followed by addition of Et.sub.3N
(0.021 ml, 0.151 mmol). The resulting suspension was stirred at
room temperature for two hours, and concentrated. It was
re-dissolved in 3 mL of THF. To this solution was added n-butyl
amine (0.014 mL, 0.139 mmol), followed by Et.sub.3N (0.021 ml,
0.151 mmol). The mixture was stirred at room temperature for two
hours, concentrated, and purified by preparative TLC plate eluting
with EtOAc:CH.sub.2Cl.sub.2:MeOH (1:1:0.05) to provide 14 mg
off-white solid as the title compound. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.65 Hz), 7.72 (s, 1H), 7.16 (s,
4H), 6.65 (d, 1H, J=5.47 Hz), 4.65 4.54 (m, 2H), 4.23 4.11(m, 2H),
3.14 3.05(m, 2H), 2.39 2.33 (m, 2H), 1.53 1.40(m, 2H), 1.39 1.27(m,
2H), 0.92 0.83(m, 3H). LCMS (ESI+) [M+H]/z Calc'd 426, found 426.
Anal. (C.sub.22H.sub.23N.sub.3O.sub.4S) C, H, N.
Example 87
N-(5-Chloro-pyridin-2-yl)-2-[4-(7-methoxy-quinolin-4-yloxy)-phenyl]-acetam-
ide
##STR00161## A mixture of 3-methoxyaniline (25 g, 204 mmol) A and
diethyl (ethoxymethylene) malonate (44 g, 204 mmol) B were heated
in an oil bath to 150.degree. C. for 40 minutes. EtOH was generated
when the temperature reached 132.degree. C. and collected. The
reaction flask was moved away from oil bath and phenyl ether (70
mL) was added into the reaction mixture. The oil bath was preheated
to 270.degree. C. The reaction was heated at 270.degree. C. (oil
bath temperature) for 15 minutes. The reaction mixture was poured
slowly into 800 ml of hexane with stirring. Ethyl
4-hydroxy-7-methoxyquinoline-3-carboxylate C was precipitated,
filtrated, washed with hexane and dried (28.4 g, 56% yield).
A solution of compound C (4.2 g) and KOH (3 g, 3 eq.) in 40 mL of
EtOH/H.sub.2O (1:1) was heated by microwave to 180.degree. C. for
50 minutes. The mixture was cooled to room temperature, poured into
water (100 mL), neutralized with AcOH to pH 7 and saturated with
NaCl. The solution was extracted with THF (3.times.300 mL) and
concentrated to yield 3.1 g of 7-methoxyquinolin-4-ol D as a
solid.
Compound D (7.4 g) was dissolved in 20 mL of POCl.sub.3. The
solution was heated to reflux for 2 hours. The excess amount of
POCl.sub.3 was removed by evaporation under vacuum. The residue was
neutralized with NH.sub.4OH to pH .about.7 and extracted with
EtOAc. The organic layer was concentrated and purified by
chromatography on a silica gel column using hexane/ethylacetate
(3:1) to give 6.5 g of 4-chloro-7-methoxyquinoline as E as a yellow
solid.
Intermediate 87a: [4-(7-Methoxy-quinolin-4-yloxy)-phenyl]-acetic
acid
##STR00162## A mixture of 4-chloro-7-methoxy-quinoline (200 mg,
1.036 mmol), 4-hydroxyphenylacetic acid (158 mg, 1.036 mmol), and
Cs.sub.2CO.sub.3 (1.02 g, 3.11 mmol) in 2 mL of DMSO was heated at
100.degree. C. overnight. The mixture was then cooled to room
temperature. EtOAc and water were added. The aqueous layer was
acidified with 1 N HCl until a precipitate was formed. The solid
was filtered and washed with water. The solid was dried in a
vacuum-oven at 60.degree. C. overnight. The intermediate 87a (160
mg) was obtained as a brown solid. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.63 (s, 1H), 8.39 (d, 1H, J=9.04 Hz), 7.47
7.28 (m, 4H), 7.16 (d, 2H, J=7.72 Hz), 6.76 (s, 1H), 3.93 (s, 3H),
3.58(s, 2H). LCMS (ESI+) [M+H]/z Calc'd 310, found 310.
The compound of Example 87 was prepared from intermediate 87a and
2-amino-5-chloro pyridine following Method A. .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 8.42 (d, 1H, J=5.47 Hz), 8.20 (s, 1H), 8.18 (d,
1H, J=6.97 Hz), 8.05 (d, 1H, J=8.86 Hz), 7.71 7.65 (m, 1H), 7.43
(d, 2H, J=8.47 Hz), 7.43 (d, 1H, J=2.26 Hz), 7.22 7.10 (m, 1H),
7.12 (d, 2H, J=8.67 Hz), 6.24 (d, 1H, J=5.47 Hz), 3.89 (s, 3H),
3.73(s, 2H). LCMS (ESI+) [M+H]/z Calc'd 420, found 420. Anal.
(C.sub.23H.sub.18N.sub.3O.sub.3Cl.0.2CH.sub.2Cl.sub.2.0.5MeOH) C,
H, N.
Biological Testing--Enzyme Assays
The stimulation of cell proliferation by growth factors such as
VEFG, FGF, and others is dependent upon their induction of
autophosphorylation of each of their respective receptor's tyrosine
kinases. Therefore, the ability of a protein kinase inhibitor to
block cellular proliferation induced by these growth factors is
directly correlated with its ability to block receptor
autophosphorylation. To measure the protein kinase inhibition
activity of the compounds, the following constructs were
devised.
(i) VEGF-R2 Construct for Assay:
This construct determines the ability of a test compound to inhibit
tyrosine kinase activity. A construct (VEGF-R2D50) of the cytosolic
domain of human vascular endothelial growth factor receptor 2
(VEGF-R2) lacking the 50 central residues of the 68 residues of the
kinase insert domain was expressed in a baculovirus/insect cell
system. Of the 1356 residues of full-length VEGF-R2, VEGF-R2D50
contains residues 806 939 and 990 1171, and also one point mutation
(E990V) within the kinase insert domain relative to wild-type
VEGF-R2. Autophosphorylation of the purified construct was
performed by incubation of the enzyme at a concentration of 4 mM in
the presence of 3 mM ATP and 40 mM MgCl.sub.2 in 100 mM HEPES, pH
7.5, containing 5% glycerol and 5 mM DTT, at 4.degree. C. for 2 h.
After autophosphorylation, this construct has been shown to possess
catalytic activity essentially equivalent to the wild-type
autophosphorylated kinase domain construct. See Parast et al.,
Biochemistry, 37, 16788 16801 (1998).
(ii) FGF-R1 Construct for Assay:
The intracellular kinase domain of human FGF-R1 was expressed using
the baculovirus vector expression system starting from the
endogenous methionine residue 456 to glutamate 766, according to
the residue numbering system of Mohammadi et al., Mol. Cell. Biol.,
16, 977 989 (1996). In addition, the construct also has the
following 3 amino acid substitutions: L457V, C488A, and C584S.
Example A
VEGF-R2 Assay: Coupled Spectrophotometric (FLVK-P) Assay
The production of ADP from ATP that accompanies phosphoryl transfer
was coupled to oxidation of NADH using phosphoenolpyruvate (PEP)
and a system having pyruvate kinase (PK) and lactic dehydrogenase
(LDH). The oxidation of NADH was monitored by following the
decrease of absorbance at 340 nm (e.sub.340=6.22 cm.sup.-1
mM.sup.-1) using a Beckman DU 650 spectrophotometer. Assay
conditions for phosphorylated VEGF-R2D50 (indicated as FLVK-P in
the tables below) were the following: 1 mM PEP; 250 mM NADH; 50
units of LDH/mL; 20 units of PK/mL; 5 mM DTT; 5.1 mM
poly(E.sub.4Y.sub.1); 1 mM ATP; and 25 mM MgCl.sub.2 in 200 mM
HEPES, pH 7.5. Assay conditions for unphosphorylated VEGF-R2D50
(indicated as FLVK in the tables) were the following: 1 mM PEP; 250
mM NADH; 50 units of LDH/mL; 20 units of PK/mL; 5 mM DTT; 20 mM
poly(E.sub.4Y.sub.1); 3 mM ATP; and 60 mM MgCl.sub.2 and 2 mM
MnCl.sub.2 in 200 mM HEPES, pH 7.5. Assays were initiated with 5 to
40 nM of enzyme. K.sub.i values were determined by measuring enzyme
activity in the presence of varying concentrations of test
compounds. The percent inhibition at 50 nm (% inhibition@50 nm) was
determined by linear least-squares regression analysis of
absorpbance as a function of time. The binding inhibitions were
fitted to equation as described by Morrison. The data were analyzed
using Enzyme Kinetic and Kaleidagraph software.
Example B
FGF-R Assay
The spectrophotometric assay was carried out as described above for
VEGF-R2, except for the following changes in concentration:
FGF-R=50 nM, ATP=2 mM, and poly(E4Y1)=15 mM.
Example C
HUVEC+VEGF Proliferation Assay
This assay determines the ability of a test compound to inhibit the
growth factor-stimulated proliferation of human umbilical vein
endothelial cells ("HUVEC"). HUVEC cells (passage 3 4, Clonetics,
Corp.) were thawed into EGM2 culture medium (Clonetics Corp) in T75
flasks. Fresh EGM2 medium was added to the flasks 24 hours later.
Four or five days later, cells were exposed to another culture
medium (F12K medium supplemented with 10% fetal bovine serum (FBS),
60 mg/mL endothelial cell growth supplement (ECGS), and 0.1 mg/mL
heparin). Exponentially-growing HUVEC cells were used in
experiments thereafter. Ten to twelve thousand HUVEC cells were
plated in 96-well dishes in 100 ml of rich, culture medium
(described above). The cells were allowed to attach for 24 hours in
this medium. The medium was then removed by aspiration and 105 ml
of starvation media (F12K+1% FBS) was added to each well. After 24
hours, 15 ml of test agent dissolved in 1% DMSO in starvation
medium or this vehicle alone was added into each treatment well;
the final DMSO concentration was 0.1%. One hour later, 30 ml of
VEGF (30 ng/mL) in starvation media was added to all wells except
those containing untreated controls; the final VEGF concentration
was 6 ng/mL. Cellular proliferation was quantified 72 hours later
by MTT dye reduction, at which time cells were exposed for 4 hours
MTT (Promega Corp.). Dye reduction was stopped by addition of a
stop solution (Promega Corp.) and absorbance at 595 nm was
determined on a 96-well spectrophotometer plate reader.
Example D
Mouse PK Assay
The pharmacokinetics (e.g., absorption and elimination) of drugs in
mice were analyzed using the following experiment. Test compounds
were formulated as a suspension in a 30:70 (PEG 400: acidified
H.sub.2O) vehicle. This solution was administered orally (p.o.) and
intraperitoneally (i.p.) at 50 mg/kg to two distinct groups (n=4)
of B6 female mice. Blood samples were collected via an orbital
bleed at time points: 0 hour (pre-dose), 0.5 hr, 1.0 hr, 2.0 hr,
and 4.0 hr post dose. Plasma was obtained from each sample by
centrifugation at 2500 rpm for 5 min. Test compound was extracted
from the plasma by an organic protein precipitation method. For
each time bleed, 50 .mu.L of plasma was combined with 1.0 mL of
acetonitrile, vortexed for 2 min. and then spun at 4000 rpm for 15
min. to precipitate the protein and extract out the test compound.
Next, the acetonitrile supernatant (the extract containing test
compound) was poured into new test tubes and evaporated on a hot
plate (25.degree. C.) under a steam of N.sub.2 gas. To each tube
containing the dried test compound extract, 125 .mu.L of mobile
phase (60:40, 0.025 M NH.sub.4H.sub.2PO.sub.4+2.5 mL/L
TEA:acetonitrile) was added. The test compound was resuspended in
the mobile phase by vortexing and more protein was removed by
centrifugation at 4000 rpm for 5 min. Each sample was poured into
an HPLC vial for test compound analysis on an Hewlett Packard 1100
series HPLC with UV detection. From each sample, 95 .mu.L was
injected onto a Phenomenex-Prodigy reverse phase C-18,
150.times.3.2 mm column and eluted with a 45 50% acetonitrie
gradient run over 10 min. Test-compound plasma concentrations
(.mu.g/mL) were determined by a comparison to standard curve (peak
area vs. conc. .mu.g/mL) using known concentrations of test
compound extracted from plasma samples in the manner described
above. Along with the standards and unknowns, three groups (n=4) of
quality controls (0.25 .mu.g/mL, 1.5 .mu.g/mL, and 7.5 .mu.g/mL)
were run to insure the consistency of the analysis. The standard
curve had an R.sub.2>0.99 and the quality controls were all
within 10% of their expected values. The quantitated test samples
were plotted for visual display using Kalidagraph software and
their pharmacokinetic parameters were determined using WIN NONLIN
software.
Example E
Human Liver Microsome (HLM) Assay
Compound metabolism in human liver microsomes was measured by LC-MS
analytical assay procedures as follows. First, human liver
microsomes (HLM) were thawed and diluted to 5 mg/mL with cold 100
mM potassium phosphate (KPO.sub.4) buffer. Appropriate amounts of
KPO.sub.4 buffer, NADPH-regenerating solution (containing B-NADP,
glucose-6-phosphate, glucose-6-phosphate dehydrogenase, and
MgCl.sub.2), and HLM were preincubated in 13.times.100 mm glass
tubes at 37.degree. C. for 10 min. (3 tubes per test
compound--triplicate). Test compound (5 .mu.M final) was added to
each tube to initiate reaction and was mixed by gentle vortexing,
followed by incubation at 37.degree. C. At t=0, and 2 h, a 250-uL
sample was removed from each incubation tube to separate
12.times.75 mm glass tubes containing 1 mL ice-cold acetonitrile
with 0.05 .mu.M reserpine. Samples were centrifuged at 4000 rpm for
20 min. to precipitate proteins and salt (Beckman Allegra 6KR, S/N
ALK98D06, #634). Supernatant was transferred to new 12.times.75 mm
glass tubes and evaporated by Speed-Vac centrifugal vacuum
evaporator. Samples were reconstituted in 200 .mu.L 0.1% formic
acid/acetonitrile (90/10) and vortexed vigorously to dissolve. The
samples were then transferred to separate polypropylene
microcentrifuge tubes and centrifuged at 14000.times.g for 10 min.
(Fisher Micro 14, S/N M0017580). For each replicate (#1 3) at each
timepoint (0 and 2 h), an aliquot sample of each test compound was
combined into a single HPLC vial insert (6 total samples) for LC-MS
analysis, which is described below.
The combined compound samples were injected into the LC-MS system,
composed of a Hewlett-Packard HP1100 diode array HPLC and a
Micromass Quattro II triple quadruple mass spectrometer operating
in positive electrospray SIR mode (programmed to scan specifically
for the molecular ion of each test compound). Each test compound
peak was integrated at each timepoint. For each compound, peak area
at each timepoint (n=3) was averaged, and this mean peak area at 2
h was divided by the average peak area at time 0 hour to obtain the
percent test compound remaining at 2 h.
Example F
KDR (YEGFR2) Phosphorylation in PAE-KDR Cells Assay
This assay determines the ability of a test compound to inhibit the
autophosphorylation of KDR in porcine aorta endothelial (PAE)-KDR
cells. PAE cells that overexpress human KDR were used in this
assay. The cells were cultured in Ham's F12 media supplemented with
10% fetal bovine serum (FBS) and 400 ug/mL G418. Thirty thousands
cells were seeded into each well of a 96-well plate in 75 mL of
growth media and allowed to attach for 6 hours at 37.degree. C.
Cells were then exposed to the starvation media (Ham's F12 media
supplemented with 0.1% FBS) for 16 hours. After the starvation
period was over, 10 mL of test agent in 5% DMSO in starvation media
were added to the test wells and 10 mL of the vehicle (5% DMSO in
starvation media) were added into the control wells. The final DMSO
concentration in each well was 0.5%. Plates were incubated at
37.degree. C. for 1 hour and the cells were then stimulated with
500 ng/ml VEGF (commercially available from R & D System) in
the presence of 2 mM Na.sub.3VO.sub.4 for 8 minutes. The cells were
washed once with 1 mm Na.sub.3VO.sub.4 in HBSS and lysed by adding
50 mL per well of lysis buffer. One hundred mL of dilution buffer
were then added to each well and the diluted cell lysate was
transferred to a 96-well goat ant-rabbit coated plate (commercially
available from Pierce) which was pre-coated with Rabbit anti Human
Anti-flk-1 C-20 antibody (commercially available from Santa Cruz).
The plates were incubated at room temperature for 2 hours and
washed seven times with 1% Tween 20 in PBS. HRP-PY20 (commercially
available from Santa Cruz) was diluted and added to the plate for a
30-minute incubation. Plates were then washed again and TMB
peroxidase substrate (commercially available from Kirkegaard &
Perry) was added for a 10-minute incubation. One hundred mL of 0.09
N H.sub.2SO.sub.4 was added to each well of the 96-well plates to
stop the reaction. Phosphorylation status was assessed by
spectrophotometer reading at 450 nm. IC.sub.50 values were
calculated by curve fitting using a four-parameter analysis.
Example G
PAE-PDGFRb Phosphorylation in PAE-PDGFRB Cells Assay
This assay determines the ability of a test compound to inhibit the
autophosphorylation of PDGFRb in porcine aorta endothelial
(PAE)-PDGFRb cells. PAE cells that overexpress human PDGFRb were
used in this assay. The cells were cultured in Ham's F12 media
supplemented with 10% fetal bovine serum (FBS) and 400 ug/ml G418.
Twenty thousands cells were seeded in each well of a 96-well plate
in 50 mL of growth media and allowed to attach for 6 hours at
37.degree. C. Cells were then exposed to the starvation media
(Ham's F12 media supplemented with 0.1% FBS) for 16 hours. After
the starvation period was over, 10 mL of test agent in 5% DMSO in
starvation media were added to the test wells and 10 mL of the
vehicle (5% DMSO in starvation media) were added into the control
wells. The final DMSO concentration in each well was 0.5%. Plates
were incubated at 37.degree. C. for 1 hour and the cells were then
stimulated with 1 mg/mL PDGF-BB (R & D System) in the presence
of 2 mM Na.sub.3VO.sub.4 for 8 minutes. The cells were washed once
with 1 mm Na.sub.3VO.sub.4 in HBSS and lysed by adding 50 mL per
well of lysis buffer. One hundred mL of dilution buffer were then
added to each well and the diluted cell lysate was transferred to a
96-well goat ant-rabbit coated plate (Pierce), which was pre-coated
with Rabbit anti Human PDGFRb antibody (Santa Cruz). The plates
were incubated at room temperature for 2 hours and washed seven
times with 1% Tween 20 in PBS. HRP-PY20 (Santa Cruz) was diluted
and added to the plate for a 30-minute incubation. Plates were then
washed again and TMB peroxidase substrate (Kirkegaard & Perry)
was added for a 10-minute incubation. One hundred mL of 0.09 N
H.sub.2SO.sub.4 was added into each well of the 96well plate to
stop the reaction. Phosphorylation status was assessed by
spectrophotometer reading at 450 nm. IC.sub.50 values were
calculated by curve fitting using a four-parameter analysis.
The results of the testing of the compounds using various assays
are summarized in Table 1.
TABLE-US-00001 TABLE 1 FLVK Ki FGF-P (% inh @ 50 nM) HUVEC + VEGF A
= >10 nM A = 0 5% IC50 A = >10 nM B = 1 10 nM B = 5 10% B = 1
10 nM Example C = <1 nM C = >10% C = <1 nM Number NT = not
tested NT = not tested NT = not tested 1 C A NT 2 A A NT 3 NT A C 4
C B B 5 NT A NT 6 C B A 7 C B C 8 B A A 9 C* B C 10 C A C 11 C A A
12 C B A 13 C B A 14 C B B 15 C A C 16 B A A 17 C C A 18 C B A 19 B
A A 20 B A B 21 NT A B 22 NT A C 23 C A C 24 C A B 25 C B B 26 A NT
NT 27 C A B 28 C B C 29 B A A 30 C A B 31 C A C 32 B A A 33 C* A A
34 NT NT NT 35 A A NT 36 NT A A 37 NT NT NT 38 NT A A 39 NT A NT 40
A A NT 41 B B B 42 A A NT 43 NT NT NT 44 NT NT NT 45 NT NT NT 46 NT
NT NT 47 NT NT NT 48 NT NT NT 49 C C B 50 B C A 51 B A A 52 B C A
53 C A B 54 A A NT 55 C A C 56 NT A A 57 A A NT 58 A A NT 59 A A NT
60 B A A 61 B A A 62 B A A 63 A A NT 64 A A NT 65 A A NT 66 A A A
67 A A NT 68 A A NT 69 A A A 70 A B NT 71 A A NT 72 NT A NT 73 NT A
NT 74 B A A 75 A A NT 76 B A A 77 A A NT 78 A C NT 79 A A NT 80 A B
NT 81 NT NT NT 82 A A NT 83 A B NT 84 A B NT 85 NT NT NT 86 B A C
87 B A A
Examples of Pharmaceutical Formulations
The exemplary compounds described above may be formulated into
pharmaceutical compositions according to the following general
examples.
Example I
Parenteral Composition
To prepare a parenteral pharmaceutical composition suitable for
administration by injection, 100 mg of a water-soluble salt of a
compound of Formula I is dissolved in DMSO and then mixed with 10
mL of 0.9% sterile saline. The mixture is incorporated into a
dosage unit form suitable for administration by injection.
Example II
Oral Composition
To prepare a pharmaceutical composition for oral delivery, 100 mg
of a compound of Formula I is mixed with 750 mg of lactose. The
mixture is incorporated into an oral dosage unit for, such as a
hard gelatin capsule, which is suitable for oral
administration.
It is to be understood that the foregoing description is exemplary
and explanatory in nature, and is intended to illustrate the
invention and its preferred embodiments. Through routine
experimentation, the artisan will recognize apparent modifications
and variations that may be made without departing from the spirit
of the invention. Thus, the invention is intended to be defined not
by the above description, but by the following claims and their
equivalents.
* * * * *